, 2004). The previously designed approach to produce and resuscitate NC cells was applied to study M. smegmatis

strain with inactivated hlp gene. Our experiments revealed that M. smegmatisΔhlp strain and its derivatives developed in the modified Hartman-de-Bont medium (without K+) at similar growth rates compared with the wild-type strain (data not shown). At the stationary phase, the Δhlp strain entered the NC state (0 CFU) later than the Wt-pMind strain with find more the empty plasmid (90–96 vs. 68–70 h) (Fig. 1a) or wild type (not shown). Complemented strain Δhlp∷hlp harboring the hlp gene on the plasmid entered the NC state only 2 h later than the Wt-pMind strain (data not shown). Next, NC cells of Wt-pMind and Δhlp selleck compound strains

were tested for their ability to resuscitate in the presence of recombinant M. luteus RpfSm protein, which appeared to be more active and stable during storage compared with full-length M. luteus Rpf (unpublished data). Contrary to Wt-pMind, NC cells of the strain Δhlp failed to resuscitate in the liquid medium with RpfSm (Fig. 2). NC cells of the complemented strain Δhlp∷hlp were partially resuscitated by RpfSm (Fig. 2). Therefore, the lack of Hlp resulted in transition of mycobacterial cells to an irreversibly NC (or likely, moribund) state under chosen conditions (K+ depletion) but not to a resuscitatable dormancy. Taken together, these data led to the conclusion that Hlp does not affect the onset of the transition to nonculturability but tuclazepam seems to be essential for cell viability at a later stage of dormancy. It was noteworthy that the strain Δhlp∷rpf, lacking the hlp gene and harboring the plasmid-carried rpf gene, substantially differed from the Δhlp in culturability when cultivated in the modified Hartman-de-Bont medium. In particular, Δhlp∷rpf cells maintained the ability to produce colonies even during a prolonged (180 h) stationary phase, as opposed to Δhlp and Δhlp-AGH strains (Fig. 1b). However, insertion of the rpf gene to

wild-type M. smegmatis (Wt-AGR) caused the development of NC state of cells incubated in the same medium and conditions (Fig. 1b), as reported previously (Shleeva et al., 2004). To ensure that the maintenance of plateability in long-stored Δhlp∷rpf cultures was indeed due to Rpf production, we studied the Δhlp∷rpf mut strain (Table 1) with the rpf gene disrupted by site-directed mutagenesis (Mukamolova et al., 2006). Our experiments demonstrated that mutations in the rpf gene restored the ability to adopt the NC state under the given cultivation conditions (Fig. 1b). Thus, the combined action of a downshift of Hlp and an upshift of Rpf greatly prolonged the ability of M. smegmatis to endure starvation in a completely culturable state, revealing opposite effects of two proteins on the formation of ‘nonculturability’. Earlier we found that M.

, 2009). IIANtr is a component of the Ntr-phosphotransferase system that is conserved in many proteobacteria (Deutscher et al., 2006). In this system, EINtr (encoded by ptsP) transfers phosphoryl groups to NPr, which subsequently phosphorylates protein IIANtr (Rabus et al., 1999; Zimmer et al., 2008). The Ntr-PTS works in parallel to the phosphoenolpyruvate : carbohydrate phosphotransferase system (transport-PTS) in E. coli. Because no obvious phosphoryl group acceptor was identified for phospho-IIANtr, it was

speculated that this system has regulatory rather than transport function. Earlier it was shown that dephosphorylated IIANtr binds and inhibits the low-affinity K+ transporter TrkA, suggesting that the Ntr-PTS somehow regulates K+ homeostasis (Lee et al., 2007). Thereupon, it was demonstrated that kdp promoter activity is stimulated

KU-60019 by the dephosphorylated form of IIANtr (Fig. 2c). The connection between the Ntr-PTS and the Kdp system was found in a search for a potential phospho-acceptor for IIANtr. Overproduction of IIANtr resulted in the accumulation of a phosphorylated protein that was identified as phospho-KdpB (Lüttmann et al., 2009). In parallel, a transposon clone library was screened for enhanced expression of kdpFABC in the presence of 5 mM K+, a concentration that normally represses the expression of the target operon. Two independent transposon mutants were isolated out of 9000 tested, in which kdpFABC expression was significantly elevated. Both insertions

mapped in gene ptsP that encodes EINtr, providing independent evidence for the inter-relation between Kdp and Ntr-PTS (Lüttmann et al., 2009). Two-hybrid data and biochemical selleck compound analysis revealed Thymidine kinase that the nonphosphorylated IIANtr interacts with KdpD and stimulates its autokinase activity. Subsequently, the level of phospho-KdpE is increased and kdpFABC is induced. It was also found that the supplied carbon source influenced kdpFABC expression, which might be related to a ‘cross-talk’ between the Ntr-PTS and the transport-PTS. When cells utilized preferred carbohydrates such as glucose, which results in dephosphorylation of the transport-PTS and also of IIANtr, kdpFABC expression was enhanced. In summary, the Ntr-PTS links carbohydrate metabolism and K+ homeostasis via the KdpD/KdpE system. Based on the results described above, the following model for activation, internal signaling, and signal integration of the KdpD/KdpE system is proposed (Fig. 2). The histidine kinase KdpD perceives different chemical stimuli from the cytoplasm (K+ concentration, ionic strength, and ATP content) that modulate the ratio between kinase and phosphatase activity. Upon activation, KdpD is transferred into the ‘ON’ state that is characterized by a high kinase to phosphatase ratio. The transition between ‘OFF’ and the ‘ON’ involves alterations of electrostatic interactions within KdpD. KdpD is a dimer, and autophosphorylation occurs in trans.

Xylella fastidiosa may use gene-regulatory mechanisms to respond to changing environments within the xylem of plants, and host range may

in part be determined by differential regulation of virulence genes in different host xylem environments. CHIR-99021 research buy Host plant resistance has been recognized as the most cost-effective and environmentally safe method for controlling many major microbial pathogens of economic plants. Understanding the underlying biochemical mechanisms of host resistance may lead to the development of resistant varieties or anti-X. fastidiosa chemicals useful in preventing disease in established grapevine. Identification of specific chemical components of citrus xylem fluid that influence the expression of virulence genes in X. fastidiosa

is underway. This work was supported in part by the University of California’s Pierce’s Disease Research Grants Program via a grant from USDA CSREES, the California Department of Food and Agriculture Pierce’s Disease/Glassy-winged Sharpshooter Board, and the University of California Agricultural Experiment Station. “
“The nasST operon encodes the transcriptional regulators of assimilatory nitrate reductase operons in phylogenetically diverse bacteria. NasT is a RNA-binding antiterminator and helps RNA polymerase read through the regulatory terminator sequences upstream of the structural genes. NasS senses nitrate and nitrite and regulates the activity of NasT through stoichiometric interaction. In this study, we analyzed the Selleckchem RG7422 nasST sequence in Azotobacter vinelandii and revealed that the nasS and nasT genes overlap by 19 nucleotides. Our genetic analyses suggested that translational initiation of NasT was coupled with NasS translation, a regulatory mechanism

that prevents overproduction GABA Receptor of NasT. The significance of tight control of nasT expression was demonstrated in a nasT-overexpression strain, where expression of the assimilatory nitrate reductase operon was deregulated. “
“The transport of organophosphates across the cytoplasma membrane is mediated by organophosphate:phosphate antiporter proteins. In this work, we present the application of a recombinant phosphoenolpyruvate:phosphate antiporter for isotopic labeling experiments in E. coli strains. The antiporters UhpT, UhpT-D388C, and PgtP were investigated regarding transport activity and growth on phosphoenolpyruvate as sole carbon source. The expression of the protein variant UhpT-D388C in a shikimic acid producing E. coli strain was used to show the successful isotopic labeling of shikimic acid from extracellular phosphoenolpyruvate. The results demonstrate the possibility of a direct incorporation of exogenously applicated glycolysis intermediates into E. coli cells for 13C-labeling experiments. “
“We have characterized swarming motility in Rhizobium leguminosarum strains 3841 and VF39SM.

Xylella fastidiosa may use gene-regulatory mechanisms to respond to changing environments within the xylem of plants, and host range may

in part be determined by differential regulation of virulence genes in different host xylem environments. learn more Host plant resistance has been recognized as the most cost-effective and environmentally safe method for controlling many major microbial pathogens of economic plants. Understanding the underlying biochemical mechanisms of host resistance may lead to the development of resistant varieties or anti-X. fastidiosa chemicals useful in preventing disease in established grapevine. Identification of specific chemical components of citrus xylem fluid that influence the expression of virulence genes in X. fastidiosa

is underway. This work was supported in part by the University of California’s Pierce’s Disease Research Grants Program via a grant from USDA CSREES, the California Department of Food and Agriculture Pierce’s Disease/Glassy-winged Sharpshooter Board, and the University of California Agricultural Experiment Station. “
“The nasST operon encodes the transcriptional regulators of assimilatory nitrate reductase operons in phylogenetically diverse bacteria. NasT is a RNA-binding antiterminator and helps RNA polymerase read through the regulatory terminator sequences upstream of the structural genes. NasS senses nitrate and nitrite and regulates the activity of NasT through stoichiometric interaction. In this study, we analyzed the find more nasST sequence in Azotobacter vinelandii and revealed that the nasS and nasT genes overlap by 19 nucleotides. Our genetic analyses suggested that translational initiation of NasT was coupled with NasS translation, a regulatory mechanism

that prevents overproduction Olopatadine of NasT. The significance of tight control of nasT expression was demonstrated in a nasT-overexpression strain, where expression of the assimilatory nitrate reductase operon was deregulated. “
“The transport of organophosphates across the cytoplasma membrane is mediated by organophosphate:phosphate antiporter proteins. In this work, we present the application of a recombinant phosphoenolpyruvate:phosphate antiporter for isotopic labeling experiments in E. coli strains. The antiporters UhpT, UhpT-D388C, and PgtP were investigated regarding transport activity and growth on phosphoenolpyruvate as sole carbon source. The expression of the protein variant UhpT-D388C in a shikimic acid producing E. coli strain was used to show the successful isotopic labeling of shikimic acid from extracellular phosphoenolpyruvate. The results demonstrate the possibility of a direct incorporation of exogenously applicated glycolysis intermediates into E. coli cells for 13C-labeling experiments. “
“We have characterized swarming motility in Rhizobium leguminosarum strains 3841 and VF39SM.

“
“International Journal of Paediatric Dentistry 2012; 22: 239–243 Background.  Adverse long-term general and dental health effects of cancer and cancer therapy during childhood have been reported. Aim.  To examine the association between chemotherapy before the age of 8 years and (1): microdontia; (2): hypodontia of premolars and permanent molars. Material and methods.  In The Danish Registry of Childhood Cancer (DBCR), we identified 203 children who met the following inclusion criteria: (1) age below 8 years at the start of treatment; (2) age between 12 to 18 years upon dental examination; (3)

had received chemotherapy The exclusion criterion was radiotherapy to the head and neck. A total of 150 children fulfilled the inclusion criteria. As controls, a random sample of 193 age-matched unexposed children learn more was included. Results.  Microdontia was found in a total of 88 teeth in 29 (19.3%) of the 150 children who had been exposed to chemotherapy, while none of the controls had microdontia of premolars or permanent molars

(difference: 19.3%; 95% CL: 13.5%; 26.4%). The earlier the exposure, the more frequent was microdontia. We found a total of 27 missing premolars and permanent molars in 14 (9.3%) of the exposed children and a total of 18 missing premolars and permanent molars in 8 (4.1%) of the controls (difference: 5.2%; 95% CL: −0.1%; 11.3%). Conclusion.  The present study confirms findings from find more previous studies that chemotherapy, especially in very young children, causes microdontia and hypodontia of premolars and permanent molars. “
“International Journal of Paediatric Dentistry 2012; 22: 180–190 Objective.  Xylitol studies suggest caries reductions in the order of 50%. Based on animal/microbial studies, erythritol potentially has caries-preventive properties. However, clinical Reverse transcriptase studies are required to confirm this.The aim of the study was to investigate the additional caries-preventive effect of xylitol/maltitol and erythritol/maltitol lozenges delivered at school,

relative to controls receiving comprehensive prevention, in a low-caries prevalence population. Methods.  A 4-year, cluster-randomized, double-blinded clinical trial. Five hundred and seventy-nine 10-year-old consenting subjects from 21 schools were randomly assigned to one of five groups. Four groups used the lozenges on school days, in three teacher-supervised sessions daily, over 1 or 2 years. The daily amount was 4.7 g/4.6 g for xylitol/maltitol and 4.5 g/4.2 g for erythritol/maltitol. The groups received free examinations and care in the public health centre. Four hundred and ninety-six children were analysed. The main outcome measure was dentin caries increment based on a clinical examination at 4 years since the start. The groups were compared in relation to the increment using hierarchical logistic regression to adjust for potential clustering. Results.

The experimental protocol was approved by the Centro de Ciências da Saúde (Universidade Federal do Rio de Janeiro) ethical committee for animal experimentation. Macrophages (1 × 105 cells) and C. parapsilosis (1 × 106 cells) were left in contact for 1 h at 37 °C in a macrophage–yeast ratio of 1 : 10 in RPMI 1640 medium pH 8.0, with the addition or not of adenosine or 5′-AMP as indicated in the legends. Coverslips were collected after this time, rinsed in phosphate-buffered saline, fixed

in Bouin’s fixative and stained with Giemsa. The percentages of infected macrophages were determined C59 wnt by counting 200 cells on triplicate coverslips of each preparation; each experiment was repeated at least three times. The association index between C. parapsilosis JAK inhibitor and macrophage cells was determined using a microscope at a magnification of × 1000 (Zeiss Axioplan 2, Germany). Representative images were taken at a magnification of × 400. The interaction between C. parapsilosis and macrophages was considered as the percentage of infected macrophages, as well as the mean number of yeast cells

per macrophage. All experiments were performed in triplicate, with similar results obtained in at least three separate cell suspensions. Statistical significance for enzymatic assays was determined by a t-test. For interaction, one-way anova and Tukey post-test were applied. P-values <0.05 were considered statistically significant. The time course of ecto-5′-nucleotidase activity on the C. parapsilosis surface was linear for 1 h and directly proportional to the number of cells (data not shown). At a pH of 4.5, intact cells were able to hydrolyze 5′-AMP at a rate of 52.44 ± 7.01 nmol Pi h−1 10−7 cells. To confirm the ectolocalization of C. parapsilosis ecto-5′-nucleotidase activity and to rule out the possibility that the observed 5′-AMP hydrolysis was the result of secreted soluble enzymes, a reaction mixture with cells was prepared and incubated in the absence of the substrate 5′-AMP. Subsequently, the suspension was centrifuged to remove the cells, and the supernatant

was checked for nucleotidase activity. The rate Fossariinae of 5′-AMP hydrolysis observed from the supernatant was <20% of that observed in intact cells (Fig. 1). In different cells, 5′-AMP is the substrate hydrolyzed by 5′-nucleotidases at the highest rates (Zimmermann, 1992; Hunsucker et al., 2005; Sträter, 2006). Intact cells of C. parapsilosis were able to hydrolyze all substrate monophosphates tested (UMP, IMP, CMP and GMP), except 3′-AMP. 5′-UMP and 5′-IMP were hydrolyzed at similar rates to that of AMP, whereas 5′-GMP and 5′-CMP presented lower rates of hydrolysis (Fig. 2). Although ecto-5′-nucleotidase activity is independent of cations (Zimmermann, 1992; Hunsucker et al., 2005), it can be modulated by the addition of Mg2+, Mn2+ or Ca2+ (Tasca et al., 2003; Borges et al., 2007).

Conclusions  The pharmacist-driven osteoporosis pathway at The Queen Elizabeth Hospital

has sustained the rate of prescription for osteoporosis therapy over a prolonged period of time. “
“The aims of the study were to (i) quantify the sales of over-the-counter (OTC) ophthalmic chloramphenicol from all community pharmacies in Wales and investigate the impact on primary care prescriptions up to 5 years after reclassification and (ii) Vincristine investigate the temporal relationship between items supplied OTC and on NHS primary care prescriptions. Primary care prescription data (2004–2010) and OTC sales data (2005–2010) for ophthalmic chloramphenicol were obtained. The quantity sold OTC was calculated from pharmacy wholesale records and sales data from a large pharmacy multiple. Spearman’s rank correlation for prescription and OTC supplies of ophthalmic

chloramphenicol was calculated for data from January 2008 to December 2010. OTC supply of chloramphenicol eye drops and ointment were both highest in 2007–2008 and represented 68% (57 708/84 304) and 48% (22 875/47 192) of the corresponding prescription volume, respectively. OSI 744 There was a steady year-on-year increase in the combined supply of OTC ophthalmic chloramphenicol and that dispensed on prescription from 144 367 items in 2004–2005 to 210 589 in 2007–2008 before stabilising in 2008–2009 and 2009–2010. A significant positive correlation was observed between prescription items and OTC sales of chloramphenicol eye drops and ointment combined (r = 0.7, P < 0.001). OTC availability increased the total quantity of ophthalmic chloramphenicol supplied in primary care compared to that seen prior to reclassification. Although growth in the sales of ophthalmic chloramphenicol OTC has stabilised and the supply pattern mirrors primary care prescribers, further work is required to investigate whether use is appropriate and whether the publication of updated practice guidance has changed this. There are three categories for human medicines in the UK, namely MRIP prescription-only medicines (POMs),

pharmacy-only (P) medicines and general sales list (GSL) medicines. POMs are only available on prescription whereas P medicines can be sold from a pharmacy under the supervision of a pharmacist. In contrast, GSL medicines can be sold from most retail outlets.[1, 2] Over-the-counter (OTC) medicines is a collective term used to describe P and/or GSL medicines that can be purchased without a prescription, although in this paper it is used exclusively to indicate supply from a community pharmacy. The main determinant of a medicine’s legal status is its safety, although factors such as side effects, monitoring requirements, route of administration, liability to misuse and risk to human health are also considered.[2] When a medicine is ‘switched’ from one legal category to another this is termed reclassification.

Impairment in wzm, noeL, and noeJ leads to defective LPS in strain Sp7. In wzm and noeJ mutants, only low-molecular-weight LPS bands were observed (Lerner et al., 2009b, c), while no LPS band was observed for the noeL mutant (Lerner et al., 2009b). In addition, substantial changes in the profile of OMPs were observed for noeJ, noeL, and wzm mutants in comparison with the wild type by SDS-PAGE (Lerner et al., 2009b, c). These mutants also showed deficient survival to salt, heat, and osmotic stresses; however, the effect of these mutations

in plant–A. brasilense interaction is still to be investigated. A recent study using atomic force microscopy revealed distinct morphological properties of flocculating A. brasilense Che1 mutants,

in comparison with the wild type. Whereas wild-type cells were shown to produce a smooth mucosal extracellular matrix, flocculating Che1 Selleckchem Alectinib mutants produced distinctive extracellular fibril structures, and likely a different structure and composition of EPS (Edwards et al., 2011). Biological nitrogen fixation by azospirilla occurs in pure culture and under optimal oxygen pressure, temperature, and carbon and energy sources (Okon, 1985). Measurable nitrogen fixation activities of azospirilla in association with plants have been demonstrated many times (Okon, 1985; Spaepen et al., 2009; Bashan & de-Bashan, 2010). However, extensive quantitative measurements of nitrogen fixation in greenhouse and field experiments as well as characterization of nitrogen fixation mutants showed that contribution of fixed nitrogen by Dabrafenib price A. brasilense does not play a major role in plant growth promotion in most systems evaluated so far (Okon, 1985; Spaepen et al., 2009). Nevertheless, nitrogen fixation ability is considered a positive attribute for rhizosphere competence of azospirilla (Okon, 1985). The two primary environmental modulators of nitrogenase synthesis and activity in A. brasilense are ammonium ions () and oxygen (O2) (Pedrosa & Elmerich, 2007; Cassan & Garcia de Salamone, 2008). The nitrogenase complex is

sensitive to oxygen and, as mentioned, carotenoids are thought to play an important role in protection against oxidative damage in A. brasilense (Hartmann & Hurek, 1988; Baldani et al., 2005). Transcription of the nitrogen fixation (nif) Staurosporine solubility dmso genes in proteobacterial diazotrophs is generally activated by the NifA protein. In many nitrogen-fixing bacteria, the nifA promoter is under control of the general nitrogen regulation (Ntr) system through the direct action of the transcriptional activator NtrC (Pedrosa & Elmerich, 2007). In contrast, in A. brasilense, NtrC is not involved in direct activation of the nifA promoter (Pedrosa & Elmerich, 2007). The activity of the NifA protein in A. brasilense is controlled by a signal transduction protein of the PII family in response to fluctuations in levels.

g., Flood et al., 1987; Turner & Deupree, 1991; Flood, 1993), and alterations in dendritic spines are region-specific, and will be discussed in terms of synapse number below. In rodents, there is loss of axospinous synapses from the layer 2 medial entorhinal cortex projection to granule cells (Geinisman selleck compound et al., 1992) and reduced synaptophysin staining in the dendritic region of CA3 pyramidal cells (Smith et al., 2000) during aging. The synaptic input to CA1 pyramidal cells from CA3, however,

does not show synapse reduction (Geinisman et al., 2004). However, a subset of the synaptic contacts in this region exhibit reduced postsynaptic density size (Nicholson et al., 2004), and electrophysiological evidence suggests that this group of synapses may reflect nonfunctional ‘silent’ synapses (Barnes et al., 1997; Burke & Barnes, 2010). Clearly, anatomical changes do occur within the hippocampus in normal aging, although they are rather subtle compared with those known to occur in pathological conditions that arise during aging, such as AD (e.g., Ballard et al., 2011). The impact

that these neurological changes have on plasticity and circuit function is discussed below. Hippocampal cell function in aging animals is strikingly well preserved. In rats it is possible to study the detailed biophysics of individual hippocampal principal cells using in vitro recording methods. Most biophysical properties in these aging cells do not change Tacrolimus (for reviews, Burke & Barnes, 2006; Hoang et al., 2012), with a small number of exceptions including a larger after-hyperpolarizing potential in CA1 pyramidal cells of old rats (e.g., Landfield & Pitler, 1984). This change may be due to an increased number of L-type calcium channels in old CA1 cells (e.g., Thibault & Landfield, 1996). This increase in channel eltoprazine numbers is hypothesized to lead to age-related disruption of neuronal calcium homeostasis, suggesting an interesting potential therapeutic target

(for review, Kumar et al., 2009). There are two additional electrophysiological changes that are observed in all three subregions of the hippocampus. These include reduced amplitude of the stimulation-induced cholinergic slow excitatory postsynaptic potential (Shen & Barnes, 1996), and an increase in gap junction-mediated electrotonic coupling between aged CA1 and CA3 pyramidal cells, as well as granule cells (Barnes et al., 1987). The former age-related change suggests reduced effectiveness of a modulatory input, and the latter increased electrical communication between cells. The alterations described above are consistent with both increased excitability (increased calcium conductance, increased electrotonic coupling) and decreased excitability (reduced cholinergic modulation) of old cells. Taken together the data suggest a complex set of mechanisms at play that may tend to keep overall cell function stable in the aged brain.

As all groups comprise neurotypical adults, we hypothesized equal variance between populations in order to control for differences in group size (Penny & Holmes, 2003). Common brain response irrespective of expertise was investigated using a minimum statistic conjunction (Nichols et al., 2005) between the three groups. Brain response specific of each group was assessed by masking exclusively the effect of this group by a global null conjunction (P < 0.05 uncorrected) of the other two groups; for instance, the contrast between Acheulean and Oldowan in Naïve is exclusively masked by a conjunction of the same contrast in Trained and Expert subjects. Our procedure used exclusive masking instead of interactions, which were

not significant at the threshold used, to favour the effects within the group of interest over selleck inhibitor the reversed effects in the Buparlisib other groups, which are included in the statistics of interactions (Culham, 2006). All contrasts were thresholded at P < 0.05 FDR-corrected with an extent threshold of 20 voxels. Anatomical localization was performed using a brain atlas (Duvernoy, 1999) and, in particular for inferior frontal and parietal clusters, functional localization made use of distribution analysis of the activated voxels on the basis of probabilistic

cytoarchitectonic maps (Eickhoff et al., 2007) implemented in SPM (Eickhoff et al., 2005). For the sake of consistency, only anatomical labels are used in the tables. Thus, clusters attributed to Brodmann area (BA) 44 were labelled ‘pars opercularis’ (Amunts et al., 1999), those attributed to BA45 were labelled ‘pars triangularis’ (Amunts et al., 1999), and those attributed to BA6 were labelled ‘precentral gyrus’ (Geyer, 2003). In the parietal cortex, clusters attributed to areas PF and PG (Caspers et al., 2006) were labelled ‘inferior parietal lobule’, and those attributed to hIP1 and hIP2 (Choi et al., 2006) were labelled ‘anterior intraparietal sulcus’. While recognizing that functional localization and anatomical landmarks may not strictly overlap in individuals, these conventions were adopted in the interest of coherence in the presentation

of results. Statistical maps were rendered mafosfamide on FreeSurfer’s fsaverage pial surface with 50 inflation steps (http://surfer.nmr.mgh.harvard.edu). In order to assess the effect of Group, local activity in clusters of interest was further characterized using the SPM extension toolbox MarsBar (http://marsbar.sourceforge.net/) to extract percentage signal change in 5-mm radius volumes centred on the maximum of each cluster, then analysed with spss. Across all subject groups, the contrast of Toolmaking conditions with Control yielded activations is a series of cortical regions, including a large cluster extending from the primary visual and lateral occipital cortices to the inferior temporal cortices, intraparietal sulci, inferior parietal cortices and postcentral gyrii bilaterally.