ORF102 and ORF103 of pAL1 were amplified by PCR using Phusion™ Hot Start High-Fidelity DNA Polymerase (Finnzymes Oy, Espoo, Finland), using total DNA of A. nitroguajacolicus Rü61a [pAL1] as the template and the primer sets listed in Supporting Information, Table S1. PCR products were subsequently ligated into pMal-c2x this website or pART2malE. Competent E. coli and A. nitroguajacolicus Rü61a [pAL1] cells were generated as described by Hanahan (1983)

and Gartemann & Eichenlaub (2001), respectively. All plasmid inserts and flanking regions were verified by sequencing (GATC Biotech AG, Konstanz, Germany). For the preparation of covalent complexes of telomeric pAL1-DNA and MBP-pORF102 or MBP-pORF103, frozen cells of A. nitroguajacolicus Rü61a [pAL1, pART2malE-ORF102] or A. nitroguajacolicus Rü61a [pAL1, pART2malE-ORF103] were thawed in 20 mM Tris/HCl buffer containing 400 mM NaCl, 1 mM

EDTA (pH 7.4) (buffer A), and 1 mM phenylmethylsulfonyl fluoride. After incubation for 30 min with 2 mg mL−1 lysozyme, crude extracts containing soluble proteins were prepared by sonication, followed by centrifugation. Supernatants were applied Panobinostat in vitro to an amylose column (5 mL bed volume), equilibrated in buffer A. After washing with the same buffer, MBP fusions were eluted with buffer A containing 20 mM maltose. Eluates were loaded to a GF/C Whatman glass microfiber filter (Whatman International Ltd, Kent, UK) (Coombs & Pearson, 1978). The immobilized complexes were washed four times with 1 M NaCl and eluted with 0.5% sodium dodecyl sulfate (SDS), 0.1 M NaCl in water (Bao & Cohen, 2001). Eluted complexes were precipitated twice with 0.1 volume Digestive enzyme of 3 M sodium acetate and 2.5 volumes of ethanol. Precipitates were redissolved in sterile water. In order to identify the DNA attached to MBP-pORF102, MBP-pORF103, or both, the redissolved complexes were used as templates in PCR reactions with GoTaq® Green DNA polymerase (Promega GmbH, Mannheim, Germany). The primer pairs for amplification of terminal regions of pAL1, an internal segment of pAL1, and a region of the chromosome of A. nitroguajacolicus Rü61a

[pAL1] are listed in Table S1. For the preparation of the MBP-pORF102 fusion protein, 5 g of frozen cells of E. coli K12 ER2508 [pLysSRARE, pMal-c2x-ORF102] were thawed in buffer A. After incubation for 30 min and addition of 1 mM MgCl2 and 10 U mL–1 benzonase, crude extracts were prepared by sonication and centrifugation as described above. The eluate from subsequent amylose affinity chromatography (performed as described above) was applied on HiTrap™ Desalting columns (4 × 5 mL, GE Healthcare, Munich, Germany) equilibrated in buffer B, consisting of 50 mM Tris/HCl (pH 8.0). The desalted eluate was loaded onto a UnoQ column (6 mL bed volume, Bio-Rad Laboratories, Munich, Germany) equilibrated in the same buffer.

While B. megaterium shows significant growth, but no adherence on the G. irregulare mycelium, V. paradoxus was fast growing and formed a dense colony around hyphae after only 45 days of incubation. This member of Bulkhoderiales was reported previously as a frequently isolated species in the Glomus intraradices hyphosphere (Mansfeld-Giese et al., 2002) and was also recovered from the hyphosphere of G. mosseae (Andrade et al., 1997). The taxon was shown to promote plant growth (Schmalenberger et al., 2008). The second most often isolated species was M. ginsengisoli. This strain also showed adherence after 45 days of incubation. Kocuria rhizophila, a soil

actinomycete, showed abundant growth and adherence after 30 days of incubation, while the Sphingomonas sp. isolate showed slow growth and little adherence on hyphae. Microbacterium and Sphingomonas genera were shown to have a potential for bioremediation Compound Library nmr by degrading hydrocarbon (Harwati et al., 2007). The Pseudomonas isolate ERK inhibitor used here as a control soil bacteria was not isolated from AMF spores, but was rather recovered from a black spruce rhizosphere, an ectomycorrhizal tree species not forming associations with AMF (Filion et al., 2004). An E. coli strain was used as a non-soil bacterial control and did not show any adherence to the fungal surface. The bacterial isolates growing in close to loose association

with the AMF mycelium may play important roles in association with the mycorrhizal symbiosis. For example, certain bacterial strains could improve mineral availability for AMF and the CYTH4 plant or could be antagonistic to certain opportunistic pathogenic organisms and improve the stability of the plant–AMF association (Xavier & Germida, 2003, Rillig et al., 2005, Marulanda-Aguirre et al., 2008). However, the data presented in this study cannot be extrapolated to the natural soil because we isolated and studied only the bacteria that can grow with

hyphal exudates as the only nutrient source, but those existing in the soil and associated with AMF that may use additional nutrient sources were not included in this study. Understanding the interactions between AMF and bacteria and their biodiversity will advance our knowledge on microbial ecology in soil and therefore could have the potential to sustain modern agriculture systems with the use of AMF and associated bacterial as biofertilizers or in bioremediation. This work was supported by NSERC discovery grants to both M.S.-A. and M.H. We thank the Canada Foundation for Innovation (CFI) for microscopy facility support to M.H. We also thank Maureen Marie-Joseph for technical assistance, Dr David Morse for comments and English editing and Dr G.V. Blomberg for kindly providing fluorescent protein plasmid vectors. Table S1. Bacterial growth and attachment on Glomus irregulare hyphae over time. Movie S1.Sphingomonas sp. Movie S2.Escherichia coli 3D1.

Electrodes with extremely high- and/or low-frequency artifacts throughout the entire recording (M = 7.2 ± 3.6) were linearly interpolated using a model of the amplitude topography at the unit sphere surface based on all nonartifactual electrodes (Perrin et al., 1990). Epochs containing nonstereotyped muscular or technical artifacts were removed. An independent component analysis approach was applied

to further reduce artifacts such as eyeblinks, horizontal eye movements, or electrocardiographic activity. Independent components representing artifacts were removed from the EEG data by back-projecting all but these components (for details, see Schneider et al., 2008). Finally, all trials that still exceeded a threshold of 100 μV were rejected automatically. On average, 1.7% (range 0.3–3.1%) of all trials were removed for each NVP-BKM120 Anti-cancer Compound Library participant. Prior to the statistical analysis, outlier trials were removed from pain ratings. To this end, the mean of intensity and unpleasantness ratings was calculated over nonpainful and painful trials separately, pooled across clips. Trials in which the ratings were below or above 3 standard deviations were excluded from further analyses. Based on this criterion, 0.29% of all trials were excluded (range 0.05–0.69%). The effect of viewing needle and Q-tip clips on

stimulus ratings was investigated by subjecting intensity and unpleasantness ratings to separate anovas with the factors visual stimulation (needle prick vs. Q-tip touch) and electrical stimulation (painful vs. nonpainful). As numerous electrical stimuli (360 painful and 360 nonpainful) were administered, it may be that habituation effects influenced the present findings (Condes-Lara et al.,

1981; Babiloni et al., 2006). To examine the possible influence of habituation on the effects in intensity selleck compound and unpleasantness ratings, additional three-way anovas, including the factor time (first and last 50% of trials within each condition), were conducted. The PDR was screened and corrected for outliers in the same way as in our recent study (Höfle et al., 2012). Eye blinks and other artifacts were removed in an interval ranging from 0.2 s before to 0.2 s after blink or artifact onset. Trials were excluded from further analyses if more than 50% of sample points within a trial were artifactual. On average, 1.2% of all trials were excluded following this criterion (range 0–3.1%). For all included trials, periods containing artifacts were linearly interpolated (Siegle et al., 2008). The PDR was normalised as follows: (data−baseline)/baseline. To establish the presence of significant effects in PDRs and to define a time interval for further analyses, point-wise running t-tests between the needle prick and the Q-tip touch trials were computed. To account for alpha error accumulation in multiple testing, time intervals were defined as being significantly different if each sample point within a 0.1 s interval reached a threshold of P = 0.05.

, 1993; Vandamme et al., 1994) and sequence data (Woese

et al., 1990; Gherna & Woese, 1992) changed the family and the genus further and provided the framework for the present Doramapimod classification. Currently, strains are assigned to the genus Flavobacterium (including 71 species to date) based on fatty acid analysis, the G+C content and a number of morphological and phenotypical characteristics following the proposal of Bernardet et al. (1996) in combination with 16S rRNA gene sequence analysis (Bernardet et al., 2002; Bernardet & Bowman, 2006). Although DNA–DNA hybridizations (DDH) are the gold standard for species identification (Stackebrandt et al., 2002), these experiments are technically challenging, laborious and time consuming. Sequence analysis of 16S rRNA genes is used for prokaryotic classification (Rossello-Mora & Amann, 2001) to provide a tentative identification. It can often limit the number of DDH experiments required. Nevertheless, the 16S rRNA gene has a limited resolving power at the species level (Fox et al., 1992; Probst et al., 1998). Within the genus Flavobacterium, values

of 97.2–98.7% 16S rRNA gene sequence similarity are found between distinct Flavobacterium species (Bernardet & Bowman, 2006). As protein-encoding genes evolve faster, they are considered more appropriate for the phylogenetic analysis of closely related species. Within the genus Flavobacterium, protein-encoding genes have not yet been used for detailed phylogenetic study. The gyrB gene was found to be a successful marker for phylogenetic analysis in several groups in other phyla, for example Acinetobacter (Proteobacteria) (Yamamoto see more & Harayama, 1996) and Micromonospora (Actinobacteria) (Kasai et al., Dynein 2000), but also in the phylum Bacteroidetes in the genus Marinilabilia and related taxa (Suzuki et al., 1999). In these studies, phylogenetic analysis based on the gyrB gene sequences was shown to be consistent with DDH and phenotypic comparison (Yamamoto & Harayama, 1996). Suzuki et al. (2001) applied gyrB gene sequencing to study the phylogenetic

relationships of marine isolates within the phylum Bacteroidetes and included two Flavobacterium species. In addition, more gyrB sequences from Flavobacterium species are becoming available in the frame of genome projects (Duchaud et al., 2007). In a previous study of aquatic and terrestrial microbial mats in Antarctica, several Flavobacterium strains were isolated that showed a low similarity to described Flavobacterium species, based on the partial or the full 16S rRNA gene sequences (Peeters et al., submitted). In the present study, we determined the gyrB gene sequence of 33 of these new Antarctic isolates and of the type strains of related Flavobacterium species to study the diversity of our isolates in more detail and to elucidate the usefulness of gyrB as a phylogenetic marker for phylogeny in the genus Flavobacterium.

, 1993; Vandamme et al., 1994) and sequence data (Woese

et al., 1990; Gherna & Woese, 1992) changed the family and the genus further and provided the framework for the present check details classification. Currently, strains are assigned to the genus Flavobacterium (including 71 species to date) based on fatty acid analysis, the G+C content and a number of morphological and phenotypical characteristics following the proposal of Bernardet et al. (1996) in combination with 16S rRNA gene sequence analysis (Bernardet et al., 2002; Bernardet & Bowman, 2006). Although DNA–DNA hybridizations (DDH) are the gold standard for species identification (Stackebrandt et al., 2002), these experiments are technically challenging, laborious and time consuming. Sequence analysis of 16S rRNA genes is used for prokaryotic classification (Rossello-Mora & Amann, 2001) to provide a tentative identification. It can often limit the number of DDH experiments required. Nevertheless, the 16S rRNA gene has a limited resolving power at the species level (Fox et al., 1992; Probst et al., 1998). Within the genus Flavobacterium, values

of 97.2–98.7% 16S rRNA gene sequence similarity are found between distinct Flavobacterium species (Bernardet & Bowman, 2006). As protein-encoding genes evolve faster, they are considered more appropriate for the phylogenetic analysis of closely related species. Within the genus Flavobacterium, protein-encoding genes have not yet been used for detailed phylogenetic study. The gyrB gene was found to be a successful marker for phylogenetic analysis in several groups in other phyla, for example Acinetobacter (Proteobacteria) (Yamamoto Lumacaftor purchase & Harayama, 1996) and Micromonospora (Actinobacteria) (Kasai et al., Adenosine triphosphate 2000), but also in the phylum Bacteroidetes in the genus Marinilabilia and related taxa (Suzuki et al., 1999). In these studies, phylogenetic analysis based on the gyrB gene sequences was shown to be consistent with DDH and phenotypic comparison (Yamamoto & Harayama, 1996). Suzuki et al. (2001) applied gyrB gene sequencing to study the phylogenetic

relationships of marine isolates within the phylum Bacteroidetes and included two Flavobacterium species. In addition, more gyrB sequences from Flavobacterium species are becoming available in the frame of genome projects (Duchaud et al., 2007). In a previous study of aquatic and terrestrial microbial mats in Antarctica, several Flavobacterium strains were isolated that showed a low similarity to described Flavobacterium species, based on the partial or the full 16S rRNA gene sequences (Peeters et al., submitted). In the present study, we determined the gyrB gene sequence of 33 of these new Antarctic isolates and of the type strains of related Flavobacterium species to study the diversity of our isolates in more detail and to elucidate the usefulness of gyrB as a phylogenetic marker for phylogeny in the genus Flavobacterium.

Further studies have reported that attention-driven top-down control can modulate the cortical representation of a range of different stimuli, from simultaneously presented motion fields to simultaneously presented visual objects (Reddy & Kanwisher, 2006; Macevoy & Epstein, 2009; Reddy & Tsuchiya, 2010), and even conjunctions of features such as color and motion (Seymour et al., 2009; see Rissman & Wagner, 2012; Tong & Pratte, 2012 for

more exhaustive reviews). In this study, we investigated if the object category of an attended stimulus can be decoded non-invasively in real-time when stimuli from two different categories are presented simultaneously. More specifically, we examined whether a classifier trained on separately presented pictures 3-Methyladenine supplier of faces and places can be used to decode the attended

object category (face or place) when both a face and a place are presented simultaneously in the form of a composite Sirolimus ic50 picture. By presenting superimposed pictures of a face and a place, we tested if object-based attention can bias the neural patterns in face- and place-selective areas towards the attended category, and if these differentiating activity patterns can be picked up on a moment-to-moment basis by multivariate pattern analysis in a real-time fMRI setting. Such an attention-driven real-time decoding setup could form the basis for a brain–computer interface (BCI) for severely paralysed and locked-in patients. Furthermore, such a system could be used to investigate if people can be trained to enhance their attention or prolong their attentional span (Jensen et al., 2011). Previous studies have shown that pictures of faces and places invoke spatially distinct and dissociable cortical regions, namely, fusiform face area (FFA) for faces and parahippocampal place area for scenes (Puce et al., 1995; Selleckchem Neratinib Kanwisher et al., 1997; Epstein et al., 1999). More recently, however, these regions have been shown to have a more overlapping and

distributed representation than previously thought (Haxby et al., 2001; Ewbank et al., 2005; Hanson & Schmidt, 2011; Mur et al., 2012; Weiner & Grill-Spector, 2012). In light of this new view, optimal decoding of faces and places from these regions call for a multivariate decoding approach that can detect these overlapping and distributed neural patterns. Therefore, in this study, we used whole-brain data to train a classifier to predict the mental state of a subject as this approach does not rely on any prior assumptions about functional localization (Laconte et al., 2007; Anderson et al., 2011; Hollmann et al., 2011; Lee et al., 2011; Xi et al., 2011; DeBettencourt et al., 2012). Moreover, the whole-brain decoder is highly suited for real-time fMRI because it automatically identifies sparse and distributed patterns of activity that are representation-specific.

[76] IL-6 also promotes increased production of MMPs.[77] Neovascularization is dependent on EC activation, migration and proliferation. Pickens et al. introduced a novel function for IL-17 as an angiogenic mediator in RA.[78] IL-17 synergizes with TNF-α in stimulating VEGF, EGF, HGF and KGF production by synovial fibroblasts. IL-17 also acts through CXCR2-dependent pathways. Fluorouracil IL-18 is a pro-inflammatory cytokine that is elevated in synovial fluids and synovial tissues in patients with RA. In the RA joint, IL-18 can contribute to the inflammatory process by inducing leukocyte extravasation through

up-regulation of EC adhesion molecules, the release of chemokines from RA synovial fibroblasts, and directly as monocytes, lymphocytes and neutrophil chemoattractants. IL-18 up-regulates the production of key regulators of osteoclastogenesis (RANKL [receptor activator of nuclear factor kappa-B ligand], M-CSF, GM-CSF and osteoprotegerin) from FLS

and also induces ICG-001 mw the serum amyloid A protein synthesis from rheumatoid synovial cells in a dose-dependent manner.[79, 80] IL-18 can also help maintain and develop the inflammatory pannus by inducing EC migration and angiogenesis. IL-18 does this function directly by binding and activating ECs and indirectly by inducing RA synovial fibroblasts to produce angiogenic chemokines and VEGF.[81] These data support the notion that IL-18 has a unique role in inducing the secretion of angiogenic elements by synovial fibroblasts in RA.[82] IL-18 is present in RA synovial fluid in high levels, where it functions as a leukocyte chemoattractant and angiogenic mediator to effect angiogenesis by inducing the secretion Terminal deoxynucleotidyl transferase of angiogenic factors

such as SDF-1α/CXCL12, MCP-1/CCL2, and VEGF.[82, 83] In conclusion, synovial fluid IL-18 levels in RA patients are good indicators of disease activity. IL-10 potentially inhibits angiogenesis through preventing the production of angiogenic mediators such as IL-8. It can also inhibit the proliferation of ECs, which is mediated by VEGF and FGF2. Also the release of angiogenic cytokies IL-1, IL-6 and TNF-α can be inhibited by IL-10. However, IL-4 activities in angiogenesis are controversial.[41] IL-4 can modulate neovascularization through pro-angiogenic and angiostatic mediators. It can also down-regulate IL-12R expression.[84] Angiostatin can inhibit angiogenesis in collagen-induced arthritis (CIA). Albini et al. presented evidence that IL-12 with potent anti-angiogenic activity is the mediator of angiostatin activity. Also it is demonstrated that angiostatin induces IL-12 mRNA synthesis by macrophages, suggesting that these cells produce IL-12 upon angiostatin stimulation.[85] IL-12 is thought to induce a cytokine cascade with antiangiogenic effects mediated by IFN-γ and angiostatic CXCR3 chemokine ligands.

There is clear evidence that a slow ascent reduces the risk of developing high-altitude illnesses.[11, 31, 39, 40] General rules for safe acclimatization at altitudes check details above 2,500 m include (1) increasing

sleeping altitude not more than 300 to 500 m per day and (2) having a rest day for every 1,000 m altitude gain or every 2 to 3 days but also prior to and/or following a greater ascent rate than usually recommended.[3, 41, 42] Heavy exercise during the ascent or high-altitude exposure appears to facilitate the development of AMS.[24, 32] Therefore, physical activity (eg, ascends) should be performed at a low intensity to minimize the individual’s exercise stress during the acclimatization period. In this context, physically fit individuals may be prevented from AMS, because the degree of the exercise stress depends on the work load related to the individual’s fitness level. However, physical fitness per se is not protective

if excessive exertion is carried out. Faster rates of ascent in more physically fit trekkers or climbers could undermine the potential protective effect of being cardiovascularly fit. In addition, as high-altitude illnesses are predominantly metabolic problems, older slower climbers may be at lower risk than younger muscularly bulkier persons with similar medical backgrounds. Thus, the mismatch between young and fit versus older less fit travelers may at least partly explain the apparent increase in AMS and related problems in the younger climbers who try to keep up with the older less fit travelers despite suffering from Tyrosine-protein kinase BLK AMS symptoms. Regular and sufficient fluid selleck kinase inhibitor intake inhibiting hypohydration prevents AMS.[24, 43] However, Castellani and colleagues reported no significant effects of hypohydration on severity of AMS[44] and hyperhydration may even have negative effects.[45] Preacclimatization in real or simulated altitude is effective in preventing AMS, but may not always be practical [eg, paying $200 per day for the additional climb up Mount Meru (4,565 m) before climbing Mount Kilimanjaro (5,895 m)]. Preacclimatization in simulated altitude solely adapts to hypoxia, whereas preacclimatization in real high altitude

includes adaptations to the specific climate conditions of high altitude (eg, cold and wind). Additionally, it can be combined with specific training to improve mountain-sport relevant skills (eg, surefootedness or walking economy). If possible, these advantages of preacclimatization by exposure to real altitude should be taken. With regard to AMS prevention, repeated daily exposures to real high altitude above 3,000 m,[31] sleeping for 2 weeks in simulated moderate altitude,[46] or 15 repeated 4-hour exposures to 4,300 m simulated altitude[47] have been shown to be effective. In a recently published review, Burtscher and colleagues concluded that daily exposures of 1 to 4 hours at a simulated altitude of about 4,000 m, repeated for 1 to 5 weeks, appeared to initiate AMS-protective effects.

[28] We found a much greater proportion of deaths among male foreign nationals. However, this is not a measurement of mortality rate, and therefore it cannot imply that the risk of death among males is higher than that of females. A significant finding of our study was that the leading causes of death among foreign nationals less than 50 years were medical illnesses. Cardiovascular disease was the leading cause of death, accounting for approximately 35%, which is consistent with studies of travelers from Australia, Canada, the United States, and Scotland.[23-27, 29, 30] We also found malignancy deaths ranked second among selleck chemicals llc all causes of deaths, accounting

for approximately 20%. This finding differs from many previously cited studies, but it was similar to the findings of Leggat and Wilks in Australia.[27] We applied the SMRs to examine Daporinad nmr whether foreign nationals in Chiang Mai City have a higher mortality than one would expect in their home countries. Surprisingly,

we found that no matter what the choice of reference populations, the results yielded very low SMRs. All of the calculated SMRs are less than 1, indicating that the mortality risk among foreign nationals visiting Chiang Mai City did not exceed mortality risk as compared with the risk in their home countries. In other words, there was no evidence of any increased risk of death from residing in or traveling to Chiang Mai City. There medroxyprogesterone were several assumptions

and limitations in this study. First, because there is no specific death registry for foreign nationals, the administrative database was assumed to be the complete database for all foreign nationals. We also assumed that the accuracy and completeness of death registration data for foreign nationals were similar to the registration data for Thai citizens. According to Tangcharoensathien et al.’s study in 2006, the completeness of the death registration in Thailand was high with 95% completeness of registration; however, only 30% of the causes of death described in the registers matched the causes determined by the medical review.[31] These inherent limitations of the death registry may impact the accuracy of our study’s results. Second, the study was unable to determine the exact number of foreign nationals visiting Chiang Mai City and it was unable to distinguish short-term travelers from long-term travelers (stay of ≥6 m). As a result, the mortality rate of foreign nationals was unable to be determined. Finally, the mortality rates in reference populations were assumed to be constant throughout the year. This assumption may influence an accuracy of the SMR estimation. Disease exacerbation among individuals with chronic illnesses while traveling is not unexpected.

1 (data not shown). Thus, Cpn60.2 appears to be the most abundant Dabrafenib chaperonin in the cell. Among the various stresses, heat shock produced large increases (typically between 20- and 200-fold) in the expression of all the genes, except for cpn60.3. We monitored heat shock-induced expression at 5, 10, 15 and 30 min after the stress. The

levels of expression of all the genes increased steadily and peaked at 15 min postshock (Fig. 3b). Ethanol and oxidative stress showed much smaller levels of change (typically between five- and 15-fold 30 and 60 min, respectively, after shocking the cells) and oxidative stress produced no change (data not shown). These results show several differences from the expression of the equivalent genes in M. tuberculosis under the same stresses (Hu et al., 2008), in particular, in the very Proteasome purification high induction by heat shock, but this may relate to the fact that microarrays that have a poorer dynamic range than qRT-PCR were used to measure expression. We also measured the expression levels of cpn60.2, cpn60.3 and cpn10 in the strain of M. smegmatis lacking cpn60.1, and found that they were not significantly different from the wild type (data not shown). As the chaperonin level is generally regulated in response

to the level of unfolded protein present in the cell, this shows that no significant

general chaperoning capacity is lost in the absence CYTH4 of Cpn60.1, supporting the model that this protein plays a more specialized role. It is not possible from these findings to determine whether or not the Cpn60.1 and Cpn60.2 proteins form mixed complexes in the cell, but we consider this to be unlikely on the basis that we have previously shown that two chaperonin proteins from Rhizobium leguminosarum, which show a much higher primary sequence identity than do the two M. smegmatis proteins, preferentially form homo-oligomers when coexpressed (Gould et al., 2007). In M. tuberculosis, regulation of expression of the duplicated cpn60 genes has been shown to involve the repressor HrcA (Stewart et al., 2002), which is widely implicated in heat shock regulation in diverse bacteria (Zuber & Schumann, 1994), and binding sites for this protein (CIRCE sequences) have been identified upstream of both genes. Mycobacterium smegmatis contains a clear homologue to the M. tuberculosis hrcA gene (MSMEG 4505: 86/95% identity/similarity). We searched the entire M. smegmatis genome for matches to the CIRCE sequence CTAGCACTCN9GAGTGCTAG, using the programme patternsearch implemented in xbase (Chaudhuri & Pallen, 2006).