Cold-induced transcripts have a long 5′ UTR containing cold-box elements described in E. coli, B. subtilis or archeabacteria (Jiang et al., 1996; Chamot et al., 1999; Hunger et al., 2006). These cis-elements modulate the stability of cold-induced mRNAs at low temperatures (Gualerzi et al., 2003). Analysis of BC0259 5′-UTR revealed the presence of such cold-box elements, with (1) one box possibly located downstream of the +1 of transcription (thus on BC0259 mRNA) and (2) two other conserved see more sequences located upstream from the +1 of transcription. The significance of these sequences upstream of the BC0259 promoter remains to be determined. However, the role of cold boxes in

the transcriptional regulation of cold genes has already been suggested elsewhere (Fang et al., 1997; Mitta et al., 1997). The cold phenotype of the 9H2 mutant is not due to a complete defect of RNA helicase encoding gene expression, as reported previously in other species with knockout mutants (Charollais et al., 2004; Ando & Nakamura, 2006). This study clearly shows that the expression level of the BC0259 gene and consequently the amount of transcripts in the cell has a huge impact on low-temperature adaptation of B. cereus. BC0259 was expressed at a higher level (about twofold) in WT cells grown at OD600 nm=0.2 when compared with cells grown at OD600 nm=1.0.

This suggests the importance of this gene during this stage of the kinetics of growth, learn more and is in agreement with the growth defect observed with the 9H2 mutant during the selleckchem lag phase. Four other RNA helicase-encoding genes are present in the B. cereus ATCC 14579 genome and may play a role in cold adaptation. Yet, they did not totally

counteract the effect of mutation in the BC0259 gene at 10 °C. The 9H2 mutant survived better than WT at a nonpermissive growth temperature, suggesting that the lower amount of BC0259 in the mutant had a positive effect on survival. Survival was improved in the presence of chloramphenicol for both the WT and the mutant, showing that a limited amount of protein synthesis was required for survival. Moreover, it has been shown that addition of chloramphenicol increases the level of cspA transcripts (Jiang et al., 1993), which is also dramatically induced in an E. coli RNA-helicase csdA mutant (Yamanaka & Inouye, 2001). This may suggest interactions between Csp and RNA helicases in B. cereus as described in B. subtilis (Hunger et al., 2006). Our transpositional approach revealed several genes that were clearly involved in low-temperature adaptation, with some also implicated in pH or salt stresses, suggesting possible cross responses in the adaptive potential of B. cereus ATCC 14579. This study also emphasizes the important role played by a DEAD-box RNA helicase in the cold-adaptive response of B. cereus, and further research is now needed to define the molecular function of this protein.

Genetic H typing confirmed the H serotype of the known strains and identified the H type of all the isolates. The numbers of strains carrying respective H types are: 10, H16; 4, H39 and 1, H45 (Table 1). There were eight O157:H16 strains, six from water in Maryland and two from ground meats in France that had identical traits, including the ɛ-eae allele (Table 1). The 15 eae-positive strains were subjected to molecular subtyping. The eight ɛ- and two β-eae-bearing O157:H16 strains shared ∼90% similarity in PFGE profiles, which were distinct from those of other

eae-carrying O157 strains (Fig. 1). The profile of the O157:H45 strain that carried α-eae shared little similarity to the other O157:non-H7 strains. Similarly, some diversity was also observed among the four κ/δ-eae-positive Akt inhibition O157:H39 strains, except for strains 7797 and 7798, which shared ∼90% profile similarity OSI-744 in vitro (Fig. 1). There were four other eae-negative O157:H16 strains, but, because this was the predominant serotype among the isolates examined, they were also included in the subtyping studies. The PFGE profiles of the eae-positive O157:H16 strains shared only ∼70% similarity to the four strains that did not carry eae (Fig. 2). Interestingly, the profiles of the six ɛ-bearing O157:H16 strains from water in Maryland

shared ∼90% similarity to one of the ɛ-bearing O157:H16 strains isolated from ground meats in France (Fig. 2). Analysis by MLST showed that the α-eae-bearing O157:H45 strain had ST-14 and the four κ/δ-bearing O157:H39 strains were ST-534, ST-563 or a new Suplatast tosilate ST that was a variant of ST-563. The eight ɛ-eae and two β-eae-positive O157:H16 strains all had ST-171, while the four eae-negative O157:H16 strains were either ST-344 or had new ST that are variants of ST-344 (Table 1). Using the MLST data, we examined the clonal relationship

between these O157:non-H7 strains, the pathogenic O157:H7 serotype and other reference EHEC, EPEC and Shigella groups. The neighbor-joining tree showed that the O157:H16 strains, including the eae-negative strains, clustered together and that the eight ɛ-eae- and two β-eae-positive strains are very closely related, if not identical (Fig. 3). All O157:H16 strains, however, are very distant to the prototypic O157:H7 strains that are in the EHEC 1 clonal group. Similarly, the other eae-positive O157:non-H7 strains were not related to the EHEC clonal groups, but instead clustered, not closely, with the EPEC clonal groups. Although strains of the O157:non-H7 serotype do not usually carry virulence genes, we examined several strains isolated from different sources and geographical areas worldwide and found that 15/57 strains of different H types carried various eae alleles. The eae gene is located on the Locus for Enterocyte and Effacement (LEE) pathogenicity island that is found mostly in EPEC and EHEC strains.

This situation is different from the United States and other European countries where the majority of advice and prescriptions are given by nurses.26–28 But training appears to be the most important key for success: training in travel medicine has indeed been associated with an improved quality of travel health advice in a number of studies, but this was independent

of whether a physician or a nurse was providing the advice.12 Second, all physicians were asked to use a computerized decision support system to help their prescriptions. Use of standardized, regularly updated and readily available sources of information on travel advice is indeed likely to improve the quality of advice. Computerized databases such as Edisan are advantageous because they incorporate detailed information, especially when there is a large this website intra-national variation in travel health risks. Third, all physicians and nurses from our travel medicine and vaccination center were aware of the study objectives and of its timelines. It remains to be seen however if similar results could be obtained in a different

setting, and could Maraviroc be sustained overtime. Despite these good results our study has some limitations. The study is monocentric and evaluation has not been performed by independent experts. We only looked at three travel-associated diseases to the detriment of other important health travel topics, and therefore we were not able to assess the quality of travel health advice in general. Nevertheless, these three important diseases Tyrosine-protein kinase BLK appear relevant for evaluation of the performance of a travel health clinic. Also, for each of these diseases we only assessed the adequacy of prescriptions to national guidelines and not the overall efficacy of the advice since we did not collect data from the same patients following their trip abroad. Indeed, in at least one case, a patient came back to us with Plasmodium falciparum malaria after being wrongly advised for malaria prophylaxis. Furthermore, although malaria prophylaxis was in accordance with national guidelines in 95.1%

of cases, the prescription of mosquito nets, another important prophylactic tool, was prescribed to only 77.7% of travelers to malaria-endemic areas.5 Finally, our results do not take in account overprescriptions of malaria prophylaxis or yellow fever vaccinations which occurred in four patients, and which should be avoided due to the cost and adverse events associated with these prescriptions, in particular the risk of vaccine-associated neurotropic or viscerotropic disease.29–31 In conclusion, appropriate advice for malaria prophylaxis, yellow fever, and hepatitis A vaccinations was provided in our travel medicine and vaccine center over a 3-month period. These good results were obtained by trained physicians in travel medicine who used a computerized decision support system. Even in this setting however, errors did occur.

Gerbella et al. (2010) Cereb. Cortex, 20, 141–168], and compared them with those to area 8/FEF (frontal eye field). Both areas

45A and 45B were the targets of highly predominant projections from the mediodorsal nucleus (MD) and of additional projections, mostly from the magnocellular check details ventral anterior and the medial pulvinar nucleus. The projection profiles from different MD subdivisions clearly distinguished these two areas from one another and from area 8/FEF. Area 45A was the target of predominant projections from parvicellular MD and of minor, albeit robust, projections from magnocellular MD. The opposite was true for area 45B: magnocellular MD was the major source of projections and parvicellular MD contributed minor, albeit robust, projections. Furthermore, area 45B, but not area 45A, was targeted by robust projections from multiform MD, the principal thalamic nucleus for area 8/FEF. These results provide further evidence for the distinctiveness of areas 45A and 45B, and support the Natural Product Library idea that area 45B is affiliated with the frontal oculomotor system, challenging the proposed homology of this area with part of the human language-related area 45 (rostral part of Broca’s region). Furthermore, the present data provide evidence for potentially robust trans-thalamic (via magnocellular MD) afferent, as well as direct and reciprocal, amygdaloid

connections of areas 45A and 45B, suggesting the contribution of emotional information to the differential role of these two areas in non-spatial information processing. “
“GABAergic transmission regulates adult neurogenesis by exerting negative feedback on cell proliferation and enabling dendrite formation and outgrowth. Further, GABAergic Phloretin synapses target differentiating dentate gyrus granule cells prior to formation of glutamatergic connections. GABAA receptors (GABAARs) mediating tonic (extrasynaptic) and phasic (synaptic) transmission are molecularly and functionally distinct, but their specific role in regulating adult neurogenesis is unknown. Using global and single-cell targeted gene deletion

of subunits contributing to the assembly of GABAARs mediating tonic (α4, δ) or phasic (α2) GABAergic transmission, we demonstrate here in the dentate gyrus of adult mice that GABAARs containing α4, but not δ, subunits mediate GABAergic effects on cell proliferation, initial migration and early dendritic development. In contrast, α2-GABAARs cell-autonomously signal to control positioning of newborn neurons and regulate late maturation of their dendritic tree. In particular, we observed pruning of distal dendrites in immature granule cells lacking the α2 subunit. This alteration could be prevented by pharmacological inhibition of thrombospondin signaling with chronic gabapentin treatment, shown previously to reduce glutamatergic synaptogenesis.

The glycopeptide antibiotic gene clusters reported for balhimycin, chloroeremomycin, A40926, A47934 and teicoplanin do not contain Fd or FdR genes (Donadio et al., 2005). Only the biosynthetic gene cluster for the related natural product complestatin contains a single Fd gene (comK) (Chiu et al., 2001). To advance in vitro studies of the cross-linking steps in glycopeptide antibiotic biosynthesis, we present efforts to identify and characterize Fd genes in the balhimycin producer A. balhimycina. The sequencing and annotation

of the entire genome is currently underway. We describe in silico analyses, which reveal 11 different Fd genes in A. balhimycina. Furthermore, we demonstrate the production and Pexidartinib price purification of two of the newly identified Fds, as well as their ability to participate in electron transfers to OxyB from both A. balhimycina and A. orientalis. The A. balhimycina DSM5908 genome was analyzed using a blast search carried out with six Fd sequences (NP-631171, NP-629284,

NP-631715, NP-625075, NP-628054 and NP-625924) from Streptomyces coelicolor A3(2) (Lamb et al., 2002; Chun et al., 2007), putidaredoxin (P00259) from Pseudomonas putida (Peterson et al., 1990; Pochapsky et al., 1994; Sevrioukova et al., 2003) and the putative ferredoxin comFd (AAK81833) from the complestatin producer Streptomyces lavendulae (Chiu et al., 2001). Eleven putative Fds, named balFd-I to balFd-XI, containing expect (E)-values

<10−6, were identified in the whole genome of A. balhimycina (Table 1). Assignment of the iron–sulfur cluster type was achieved through a blast search of the nonredundant Selleckchem MDV3100 database and a comparison with known Fds. The sequences of the ferredoxins balFd-I to balFd-XI have been deposited in the EMBL database under the accession numbers FN594523-FN594533. The genomic DNA of A. balhimycina DSM5908 was used as a PCR template to amplify the genes coding for the putative [3Fe–4S] ferredoxins balFd-V and balFd-VII. The primers used are shown below (restriction sites underlined): 5′-balFd-V: 5′-TAGACCATATGAAGGTTGTTGTCGACG-3′ 3′-balFd-V: 5′-ATCTACTCGAGGGCCTCTTCGAGGGC-3′ Carbohydrate 5′-balFd-VII: 5′-TAGACCATATGAAGGTCACCGTGGACG-3′ 3′-balFd-VII: 5′-ATCTACTCGAGCGCGTCCTCGCTCACCG-3 After digestion with NdeI and XhoI, the fragments were cloned between the NdeI/XhoI sites of plasmid pET22b (Novagen) for expression as C-terminal His6-tagged fusion proteins. The nucleotide sequence of each insert was confirmed by sequencing. For production in Escherichia coli Rosetta2(DE3)pLysS (balFd-V) or Origami2(DE3) (balFd-VII), terrific broth medium (400 mL) supplemented with antibiotics and FeSO4 (0.5 mM) was induced with isopropyl-β-d-thiogalactopyranoside (0.2 mM), and shaking at 30 °C for 6 h. Cell pellets in buffer-A (25 mL, 50 mM sodium phosphate pH 7.4, 300 mM KCl, 1 mM dithiothreitol, 1 mM phenylmethylsulfonyl fluoride, 1 mM benzamidine, 0.

It has been identified previously as an osmolyte in methanogenic Archaea (Robertson et al., 1990, 1992; Selleckchem Epacadostat Lai et al., 1991; Roberts, 2005) and also in three genera of Bacteria: the marine gammaproteobacterium Alteromonas luteoviolacea (Henrichs & Cuhel, 1985), several species of the actinobacterial genus Nocardiopsis (Galinski & Trüper, 1994; DasSarma & Arora, 2002) and the anoxigenic phototrophic bacterium Thiohalocapsa halophila DSM 6210T (Severin et al., 1992). Anionic solutes, such as α-glutamate and β-glutamate, have been considered

to be counterions for elevated intracellular K+ concentration (Roberts, 2005). In addition, the most prominent 13C chemical shifts (at salinities >3%) in NMR data were consistent with the accumulation of a compound that was previously unknown as a compatible solute in Bacteria, tentatively identified as NeABL according to connectivities in 2D-NMR experiments and 13C DEPT-135 data. 1H–13C HMQC data allowed determining the correlation between 1H and 13C shifts detected on corresponding spectra for such compounds (Supporting Information, Fig. S1). Other connectivities derived from a 1H–1H COSY indicated both CH2-CH(N)-CH2 (asymmetric carbon in β-position) and -NCH2-CH2- moieties as well KU-60019 as the CH3CO group (Fig. S2), which was further assigned to the ɛ-position by means of 1H–13C HMBC experiments, because a cross-peak

was detected between the proton in the ɛ-position (CH2 resonance at 3.20 p.p.m.) and the carbonyl of the acetyl group (13C shift at 176.7 p.p.m.)

(Fig. S3). ESI-MS analyses from collected fractions of a desalted aqueous cell extract showed a molecular peak at m/z 188.5 for the suspected compound (Fig. S4). Therefore, the molecular formula of C8H16N2O3 was proposed. The observed MS signals proved to be consistent with the theoretical isotope pattern of this molecule. Subsequently, the proposed structure was confirmed through its chromatographic preparation and purification (Figs S5–S8). enough Both 13C and 1H chemical shifts of the purified compound as well as its 1H–1H (COSY) connectivities (Fig. 2) were also consistent with the proposed structure corresponding to NeABL. Both isolated and type GSB strains were cultured in media with different NaCl concentrations to determine differences in the composition of major compatible solutes from 13C-NMR spectra. Although GSB species had essentially the same cocktail of compatible solutes, changes in the relative intensity of signal in NMR results suggested that different ratios among these compounds occurred in different strains and salt concentrations. Thus, anionic solutes, either l-α-glutamate or β-glutamate, would be the main organic compounds accumulated for osmotic balance at a low NaCl content (3%) (Fig. 1).

rTMS R7 58 ± 3%, P = 0.01; contralesional targets, 41 ± 15% vs. 65 ± 10%, P = 0.01) whereas it did not influence the detection of static targets (Static ipsilesional targets R7, 42 ± 5% vs. post-rTMS 48 ± 3%, P = 0.10; and contralesional post- rTMS R7, 38 ± 3% vs. post-rTMS 45 ± 12%, P = 0.56). These effects reverted to pre-rTMS values particularly for mid-central ipsilesional eccentricities (Moving 2: 45°, post-rTMS 50 ± 18% vs. rTMS R7 81 ± 19%, P = 0.24; 60°, 43 ± 19% vs. 67 ± 23%, P = 0.26; Fig. 8). Overall, the restoration of performance in Non-responders proved to be reversible once the rTMS regime ended,

which further supports the role of neurostimulation as being responsible for the maladaptive effects observed in this subset of animals. The intention of the experiment was to damage HDAC phosphorylation the homologue of the human posterior parietal cortex, known as pMS, and to later apply rTMS on the rostrally adjoining aMS cortex, which is known for its ability to adequately compensate lost function after lesion (see Fig. 1 for details on the anatomy). A comprehensive lesion analysis indicated that, for all animals, the majority of the injured cortical area was removed. Nonetheless, areas of incomplete GSI-IX cell line damage were found extending 1–3 mm rostrally in some subjects (n = 3 in Responders and n = 3 in

Non-responders), impinging into the aMS cortex (stereotaxic selleck chemicals levels A9–A11) or 1 mm caudally into the ventral posterior suprasylvian and the dorsal posterior suprasylvian regions (stereotaxic level P3; n = 2 in Responders and n = 3 in Non-responders). In addition, all 12 subjects showed very minor collateral damage to the pMS-adjacent visual areas such as primary visual area A19 and the splenial visual area, due to a minor but unpreventable diffusion of the neurotoxin. This spread appears to be consistent with other studies using the same methods (also see Rudolph & Pasternak, 1996; Huxlin et al.,

2008; Rushmore et al., 2010; Das et al., 2012; Supporting Information Figs S1 and S2). Quantification of injured area (mm2) showed no significant differences in the amount of lesion between groups, either for the medial (pMLS) or the lateral (pLLS) bank of the posterior parietal (pMS) cortex along the length of both pMS and aMS visual areas. Overall, the amount of spared tissue between Responders and Non-responders in both the injured pMS cortex (pMLS: 21 ± 8% vs. 14 ± 6%, P = 0.2; pLLS: 18 ± 6% vs. 15 ± 6%, P = 0.60) and the rTMS-stimulated aMS cortex (aMLS, 79 ± 7% vs. 58 ± 13%, P = 0.10 and aLLS, 79 ± 7% vs. 64 ± 13%, P = 0.10; data not shown in figure form) was not statistically different across groups. Responders and Non-responders also did not show significant differences in spared cortex at any specific coordinates across the rostral–caudal extent from pMS through aMS (medial bank, F4,32, P = 0.32; lateral bank, F4,32, P = 0.60).

We also expected to find a greater impact of CDSSs on prescribing outcomes than clinical outcomes and examined whether multi-faceted Ganetespib purchase interventions would have a greater impact than CDSS alone. We included English-language studies, published between 1990 and March 2009, reporting RCTs and strong quasi-experiments (non-randomised studies with comparison groups or interrupted

time-series designs with or without comparison groups). The studies had to: target pharmacists; compare the performance of the CDSSs to routine care and/or paper-based decision support; provide information that could be applied to a specific patient (e.g. provide advice to prescribe a particular drug, to monitor a drug or adjust the dose or to perform laboratory tests related to safe prescribing); generate information or advice to the pharmacist in an electronic format (however, subsequent delivery of information to physicians or patients could be in electronic or paper formats); and report data on at least one outcome relating to prescribing, clinical or patient outcomes (see Table 1). Studies were excluded if: interventions were based around hypothetical scenarios rather than actual clinical

practice, studies did not undertake statistical analyses or studies reported only cost outcomes. We searched Medline (1990–March find more 2009), PreMedline (18 March 2009), Embase (1990–March 2009), CINAHL (1990–March 2009) and PsycINFO (1990–March 2009). We combined keywords and subject headings to identify computer-based

decision support (e.g. decision support systems clinical, decision making computer assisted), medicines use (e.g. prescription Amino acid drug, drug utilization) and pharmacy or pharmacists. We also searched INSPEC (March 2009) and the Cochrane Database of Systematic Reviews (March 2009) including reviews and protocols published under the Effective Practice and Organisation of Care Group (EPOC). Finally, we hand-searched the reference lists of retrieved articles and reviews. Table 2 details the full search strategy. Two reviewers (JR, EW) evaluated independently the study titles and abstracts identified in the search. Full-text articles were retrieved if either reviewer considered a citation potentially relevant. Studies deemed eligible for review underwent data extraction by two reviewers (JR, EW). Disagreements were resolved by discussion to reach consensus. We extracted the following information from eligible studies: objectives, clinical setting (ambulatory or institutional care) and details of the decision-support intervention (e.g. system-initiated or user-initiated, multi-faceted or CDSS alone, clinical target). We classified studies as having a safety or a QUM focus. Given the lack of uniformity in relation to terminology about prompts, alerts and reminders we extracted details as they were reported in the articles.

, 2008) with Alexa Fluor 546-labeled phalloidin (Invitrogen) (1 : 200 dilution in BSP) for 1 h, washed twice with BSP and then mounted on a glass slide using mounting solution with 4′,6-diamidino-2-phenylindole. Once the slides were dry, coverslips were sealed using clear nail polish, and images were captured using green and red filters on an Olympus IX70 inverted fluorescence microscope equipped with a DP70 digital camera. The images were merged using imagej software (National Institute of Health). In our previous study (Puttamreddy et al., 2010), a mini-Tn5 transposon insertion library was constructed in E. coli O157:H7 strain EDL933

and 51 distinct genes/intergenic regions were identified to be involved either directly or indirectly BLZ945 chemical structure in biofilm formation. All of the 51 biofilm-negative mutants showed reduced biofilm formation in a quantitative biofilm assay at P<0.05. To test whether biofilms on different surfaces may require different gene products, we subjected all 51 Bnp mutants to a quantitative biofilm test on polystyrene, polypropylene, polyvinyl chloride and glass. There was no statistical difference in the quantity of biofilms produced on any of the surfaces tested (Supporting Information, Fig. S1). To analyze the Epigenetic animal study role of biofilm formation of E.

coli O157:H7 in adherence to T84 (human colonic epithelial) and HEp2 (human laryngeal epithelial) cells, wild type and Bnp mutants were tested for adherence in an in vitro adherence assay by both microscopic and quantitative analysis. We also constructed eae and esp deletion mutations as controls for potential adhesin–biofilm interactions.

Additionally, we tested a biofilm-positive transposon mutant (M1) as a control for nonspecific transposon effects on cellular adherence. The quantitative adherence assay (significance P<0.05) showed that all 51 Bnp mutants lost their adherence to both T84 and HEp2 cells after a 90-min infection when compared with wild type (Fig. 1, Table S1). This was also true for the eae and espAB deletion mutants as expected (Fig. 1, Table S1) because both of these genes have been shown previously to be important to cellular adherence (Yu & Kaper, 1992; Ebel et al., 1998). For microscopic analysis, all of the bacterial strains were transformed with a green fluorescent protein-expressing plasmid, pISM31. Selleckchem CHIR99021 Escherichia coli O157:H7 EDL933 wild type adhered to both cell types while all of the 51 Bnp mutants failed to adhere. A biofilm-positive mini-Tn5Km2 mutant of E. coli O157:H7 EDL933 (M1) showed no significant loss in either biofilm formation or adherence when compared with the wild type (Figs 2 and 3, also see Fig. S2 for a higher magnification of wild-type adherence to the two cell types). This eliminates the possibility of a nonspecific effect between mini-Tn5Km2 insertions and loss of adherence. To identify whether loss of adherence leads to a loss of biofilm phenotype, deletions in eae and espAB, both well-known adhesins (Jerse et al.

, 2001). This is why click here the conclusions

of Lange & Röder (2006) are based only on expectancy manipulation at the earlier time point, preventing investigation of the temporal course of attentional modulations. The present study manipulated relative stimulus probabilities in vision and touch independently, and maintained uncertainty throughout the trial by adding foils. This allowed us to gauge attention effects at early and late time intervals for each modality. If the hypothesis of cross-modal synergy in temporal orienting of attention holds, then one would expect faster RTs for all the stimuli presented at the overall expected time, regardless of modality prevalence (that is, for events in the primary or secondary, less likely, modality). Instead, if such synergistic effects fail to operate, then only the primary modality will be facilitated at the overall expected time points, without coupling of performance EPZ5676 in the secondary modality. In this case, one would expect an interaction between modality prevalence and temporal expectation. Moreover, performance in the secondary modality might abide by its relative temporal distribution independently of the primary modality. A total

of 29 participants volunteered for this experiment (two left-handed; 18 female; mean age 26.62 years, age range 18–36 years) in exchange for 8€ per hour. They all reported normal or corrected-to-normal vision and gave written informed consent to participation in the study, which is in accordance to the Declaration of Helsinki and approved by the ethics committee CEIC Parc de Mar (University Pompeu Fabra, Barcelona, Spain). The stimuli could be visual or tactile, and presented as single- or double-pulse stimulation. Visual stimuli consisted of the illumination of a yellow LED (0.025 cd/m2) at the centre of a black cardboard box (32.5 × 20 × 11 cm) placed at a lower frontal viewing distance of 35 cm from the participant (see Fig. 1). The single-pulse visual stimulus was a flash of Inositol monophosphatase 1 200 ms

and the double-pulse stimulus consisted of two 75-ms flashes, separated by a 50-ms gap. Tactile stimulation was presented on the index finger pad of the participant’s hand, which was placed spatially aligned underneath the LED delivering the visual stimuli (left- or right-hand stimulation was fixed within participant and counterbalanced between them). The tactile stimuli were delivered by a solenoid tapper (round tip, 8 mm; Miniature Solenoid Tapper, MSTC3-10M; M&E Solve). For single-pulse stimulation the tapper was lifted for 10 ms; double-pulse stimuli consisted of two 10-ms stimulations, separated by a 30-ms gap. The tactile stimulation did not cause any pain or annoyance to the participant.