Furthermore, voltage-sensitive dye imaging only provides informat

Furthermore, voltage-sensitive dye imaging only provides information related to the superficial dorsal neocortex, and it is likely that there are many additional targets of barrel cortex axons. The remainder of this review will focus on the anatomical connectivity of the mouse barrel cortex with specific reference to axonal output from the C2 barrel column. Anatomical connectivity can be studied by directly injecting classical tracers or viral vectors (which can also be used as anatomical tracers) into the specific brain region under investigation. Intrinsic

optical imaging provides a simple way to localize the functional representation this website of the mouse C2 whisker through the intact skull (Fig. 3A; Ferezou et al., 2006; Aronoff & Petersen, 2007; Lefort et al., 2009). By aligning the intrinsic optical signal with Maraviroc research buy the surface blood vessels, a small craniotomy can be made over the C2 whisker representation in S1 barrel cortex, enabling direct injection of anatomical tracers into the C2 barrel column (Fig. 3B). Injection of a lentivirus into the functionally identified C2 whisker representation localized by intrinsic optical imaging results in labelling of neurons located in the C2 barrel column (Fig. 3C). Intrinsic optical imaging therefore provides a reliable

map of S1, allowing anatomical Tyrosine-protein kinase BLK tracers to be reliably targeted to the C2 barrel column. If biotinylated dextran amine (BDA) is injected into the cortex, it is taken up locally by neuronal cell bodies and diffuses into their dendrites and axons (Fig. 3D). Because of the biotinylation, BDA can be readily stained, providing high contrast fluorescence images. BDA is therefore an anterograde tracer which can be used to study the axonal output of a given

brain area. However, it should be noted that BDA is also to some extent taken up by axons near the injection site (especially when it is pressure-injected), meaning that there is also some labelling of axons with cells bodies (and their axonal collaterals) located far from the injection site. Such collateral labelling complicates the interpretation of BDA-labelled tissue. Fluorogold (FG) injected into the cortex is taken up by axonal boutons and transported retrogradely to the soma. FG labelling is prominent in the cytoplasm of neuronal soma located in brain regions projecting to the injection site, and FG is therefore a useful retrograde tracer. These ‘classical’ anatomical methods are now complemented by a variety of viral vector strategies for labelling (Fig. 3E and F), which may offer higher specificity for anatomical tracing and, in addition, provide the opportunity for genetic manipulation of the transduced cells.

, 2003) It has been shown that a thyX knockout mutant of C glut

, 2003). It has been shown that a thyX knockout mutant of C. glutamicum was more sensitive to a DHFR inhibitor compared with a wild-type strain. This could be because both ThyA and ThyX contribute to the synthesis of the one-carbon unit for the

biosynthesis of thymidine in C. glutamicum. Moreover, because only the ΔthyX strain exhibited poor survival during the stationary growth phase, it has been 17-AAG cell line suggested that the expression levels of thyA and thyX differ in response to different growth conditions (Fivian-Hughes et al., 2012; Park et al., 2010). Sigma factors are components of RNA polymerases that bind to the core subunits of the enzyme and confer specificity to the process of transcription initiation by recognition of promoter sequences of genes and operons. The presence of seven putative sigma factors, including SigA and SigB, in C. glutamicum reflects the ability of the bacterium to adapt to various stress conditions (Kalinowski et al., 2003; Pátek & Nešvera, 2011). SigB is an alternative sigma factor that is not essential for exponential growth. Genome-wide transcription profiles of the wild-type and ΔsigB strain strains of C. glutamicum have shown that SigB is involved in amino Natural Product Library acid metabolism, carbon metabolism, stress defense, membrane processes,

and phosphorus metabolism (Ehira et al., 2008). Our primary interest in the present study was to measure the levels of ThyA and ThyX during growth of C. glutamicum. Western blot analysis with ThyA and ThyX antiserum suggested that both proteins were expressed, and that ThyX was maintained at the same level in both late-exponential and stationary phase cells. We also carried out Western blot analysis of total protein from the wild-type, ΔsigB, and sigB-complemented strains. Our results showed that SigB is responsible for the level of ThyX during transition into

the stationary growth phase. The bacterial strains used in this study are listed in Table 1. Escherichia Galactosylceramidase coli and C. glutamicum strains were cultured at 37 and 30 °C, respectively, in Luria–Bertani (LB) medium. Minimal medium used for C. glutamicum was mineral C. glutamicum citrate (MCGC) (Von der Osten et al., 1989) with glucose added to a final concentration of 1% (w/v). Ampicillin (100 μg mL−1), kanamycin (50 μg mL−1), and WR99210-HCl (3 μM) were added when needed. 5-Bromo-4-chloro-3-indolyl-β-d-galactopyranoside (X-gal, 3 μg mL−1) was used to monitor β-galactosidase production on plates. PCR was used to amplify the coding sequences of the thyA and thyX genes from C. glutamicum ATCC 13032. The DNA fragment corresponding to the thyA gene was amplified using primers pQETHYA1 and pQETHYA2, and the thyX gene was amplified using primers pETTHYX1 and pETTHYX2. The PCR fragments of thyA were digested with SmaI and HindIII, and then cloned into pQE82L (Qiagen), which was also digested with SmaI and HindIII, to yield plasmid pQE82L-thyA.

Pharmacological experiments indicated that PACAP triggers this an

Pharmacological experiments indicated that PACAP triggers this antiproliferative effect through the activation of both PAC1 and VPACs, and the cAMP–PKA pathway. In addition, PACAP receptor activation decreased both cyclin D1 mRNA and protein content. Altogether, the data support the hypothesis that PACAP is a cell-extrinsic regulator with multiple roles during retinal development, including the regulation of proliferation in a subpopulation of retinal progenitor cells. “

Ca2+ channels are rapidly inactivated by a mechanism that is termed Ca2+-dependent inactivation (CDI). In this study we investigated the influence of intracellular Ca2+ release on CDI of high-voltage-activated Ca2+ channels in rat thalamocortical Ku 0059436 relay neurons by combining voltage-clamp, Ca2+ imaging and immunological techniques. Double-pulse protocols revealed CDI, which depended on the length of the conditioning pulses. Caffeine caused a concentration-dependent increase in CDI that was accompanied LBH589 purchase by an increase in the duration

of Ca2+ transients. Inhibition of ryanodine receptors and endoplasmic Ca2+ pumps (by thapsigargin or cyclopiazonic acid) resulted in a reduction of CDI. In contrast, inhibition of inositol 1,4,5-tris-phosphate receptors by intracellular application of 2-aminoethoxy diphenyl borate or heparin did not influence CDI. The block of transient receptor potential channels by extracellular

application of 2-aminoethoxy diphenyl borate, however, resulted in a significant reduction of CDI. The central role of L-type Ca2+ channels was emphasized by the near-complete block of CDI by nifedipine, an effect only surpassed when Ca2+ was replaced by Ba2+ and chelated by 1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′,-tetraacetic acid (BAPTA). Trains of action potential-like Megestrol Acetate stimuli induced a strong reduction in high-voltage-activated Ca2+ current amplitude, which was significantly reduced when intracellular Ca2+ stores were made inoperative by thapsigargin or Ba2+/BAPTA. Western blotting revealed expression of L-type Ca2+ channels in thalamic and hippocampal tissue but not liver tissue. In summary, these results suggest a cross-signalling between L-type Ca2+ channels and ryanodine receptors that controls the amount of Ca2+ influx during neuronal activity. “
“Neurotrophin-3 (NT-3) is a trophic factor that is essential for the normal development and maintenance of proprioceptive sensory neurons and is widely implicated as an important modulator of synaptic function and development. We have previously found that animals lacking NT-3 have a number of structural abnormalities in peripheral nerves and skeletal muscles. Here we investigated whether haploinsufficiency-induced reduction in NT-3 resulted in impaired neuromuscular performance and synaptic function.

No comparative studies have been performed and hence there is no

No comparative studies have been performed and hence there is no optimal ‘gold-standard therapy’ (level of evidence 1B). We recommend that chemotherapy regimens should be combined with HAART therapy (level of evidence 1B). We recommend Epacadostat datasheet the addition of rituximab (level of evidence 1C). 4.6.1 Recommendations for IT prophylaxis We recommend

that patients with DLBCL, considered to have a high risk of CNS relapse, should be given CNS prophylaxis (IT and/or IV methotrexate) according to the same criteria as HIV-negative patients (level of evidence 1C). We recommend that prophylactic intrathecal chemotherapy should be offered to all patients with Burkitt lymphoma (level of evidence IB). 4.8.1 Recommendations for patients with relapsed/refractory aggressive ARL We recommend that patients deemed fit

for intensive chemotherapy PD0332991 mouse should receive a second-line chemotherapy regimen (level of evidence 1C), which may contain platinum (level of evidence 2C). We recommend that those patients responding to second-line chemotherapy (CR or PR) should be considered for HDT with ASCT (level of evidence 1C). 5 Primary central nervous system lymphoma (PCNSL) 5.4 Recommendations We recommend that all patients with PCNSL should be started on HAART if not already on it (level of evidence 1C). We recommend that patients with an adequate performance status should be treated, if possible, with high-dose methotrexate-containing chemotherapy regimen (level of evidence 1D). We recommend that whole brain radiotherapy is a useful palliative treatment modality for control of symptoms or should be considered as an alternative first-line treatment modality in those patients where the risks of toxicity from high-dose intravenous agents are considered unacceptable (level of evidence 1C). 6 Primary effusion lymphoma (PEL) 6.6 Recommendations

We suggest that first-line treatment of PEL in HIV-infected individuals includes CHOP-like regimens. No comparative studies have been performed and there is no optimal gold-standard therapy (level of evidence 2C). Patients, where possible, should be entered into clinical trials that are testing novel targeted approaches (GPP). 2-hydroxyphytanoyl-CoA lyase We recommend that chemotherapy regimens should be combined with HAART (level of evidence 1C). 7 Plasmablastic lymphoma 7.6 Recommendation We recommend that patients should receive HAART with systemic anthracycline-containing chemotherapy as first-line therapy (level of evidence 1C). 8 Cervical intraepithelial neoplasia (CIN) and cervical cancer 8.6 Key recommendations We recommend that all women newly diagnosed with HIV should have cervical surveillance performed by, or in conjunction with, the medical team managing their HIV infection (level of evidence 1B). An initial colposcopy and annual cytology should be performed if resources permit (level of evidence 2C).

3c, both clean and infected cells exhibited μ-calpain and m-calpa

3c, both clean and infected cells exhibited μ-calpain and m-calpain

activities. When normalized for protein GDC-0980 in vitro content, the calpain activity in the infected cells was slightly lower than the activity observed in the clean cells, without a significant difference between them [the calpain activity levels in the infected cells were 80±12.2% as compared with the levels in the clean cells (P>0.1; n=3)]. The NDMH lacked any calpain activity (Fig. 3c), consistent with the absence of calpain protein in the mycoplasma shown by immunoblotting (Fig. 3a). The results suggested that under the conditions used here, calpastatin was, to a large extent, separated from calpain in the zymography of the cell extracts of NDMH-infected cells, similar to the separation in the clean cells, allowing calpain caseinolytic activity. To further investigate the effects of mycoplasmal infection on calpain activation and activity, differentiated SH-SY5Y

cells were treated with Ca2+/ionomycin, as described in Materials and methods. μ-Calpain autolysis was enhanced in the Ca2+/ionomycin-treated clean cells, as shown by the appearance of the calpain 76 kDa band, compared with the control clean cells. Little calpain autolysis was observed in the Ca2+/ionomycin-treated infected cells, as shown by the low ratio of the 76 kDa band to the 80 kDa band in these cells, compared with that of the clean cells (Fig. 4a and b). These results suggest that the higher levels of calpastatin in the NDMH-infected cells inhibit Ca2+-promoted calpain activation. Fodrin is a known substrate for calpain, with a fodrin fragment of 150 kDa indicative of caspase and calpain activities, 145 kDa considered Daporinad clinical trial to be due to calpain activity and 120 kDa considered to be due to caspase activity (Wang, 2000). As shown in Fig. 4c and d, a significantly increased degradation of fodrin to 150/145 kDa fragments was observed in the Ca2+/ionomycin-treated clean cells (211±22% of the levels in the control clean cells); degradation of fodrin to 150/145 kDa bands was inhibited in the

Ca2+/ionomycin-treated infected cells (125±3% of the levels in the control clean cells) (Fig. 4c and d). The results suggest that calpain activity, promoted Oxymatrine by increased Ca2+ in the intact clean cells, is inhibited in the infected, calpastatin-overexpressing cells. Contamination of human cell cultures by mycoplasma is frequent, commonly detected in 15–35% of cell cultures, with rates reaching 65–80% in some surveys (Drexler & Uphoff, 2002). Contamination is often undetected, because the culture medium remains clear and the cellular morphological changes may not be obvious. Thus, mycoplasma-induced alterations in cell components, metabolism and regulation of various functions (Drexler & Uphoff, 2002; Rottem, 2003) may not be appreciated, unless specifically studied. Mycoplasma hyorhinis is one of the most common Mycoplasma species that contaminate various cell cultures (Drexler & Uphoff, 2002; Timenetsky et al., 2006).

All isolates presented ADA activity, although we could not establ

All isolates presented ADA activity, although we could not establish a relationship between isolate source and activity (Table S2). Herein, we described the biochemical properties of an ADA activity and two ADA-related sequences present on intact trophozoites of T. vaginalis. Cellular integrity was assessed, before and after the reactions, and the viability of the trophozoites was not affected by any of the conditions used in the assays. The influence of pH on the adenosine deamination in T. vaginalis was verified and the results demonstrated that the optimal pH for ADA activity reached at AZD2014 7.5. It is known that vaginal pH in noninfected women is approximately 4.3, but can vary from

below 4 to pH 7.5 during the menstrual cycle (Stevens-Simon et al., 1994). In agreement, previous studies demonstrated that the optimal pH values for ADA activities from the camel tick, H. dromedarii,

and from the trematode F. gigantica were also 7.5 (Mohamed, 2006; Ali, 2008). Cation exposures (2.5 mM) were able to decrease the adenosine deamination in T. vaginalis in approximately 50%. Higher concentration of calcium (5.0 mM) completely abolished the enzyme activity and the presence of EDTA, a chelating agent, restored ADA activity. GSK2118436 mw Previous data showed that zinc and other divalent cations are able to interact with other amino acid residues and induce an inhibition of the enzyme activity (Cooper et al., 1997; Mohamed, 2006; Rosemberg et al., 2008). Because zinc is toxic to Vitamin B12 T. vaginalis, we could not perform the experiments on the influence of this metal in ADA activity in intact trophozoites (Langley et al., 1987; Houang et al., 1997). Additional

studies are necessary to explain the relevance of the inhibition of ADA activity by calcium and magnesium in T. vaginalis physiology, because magnesium is the most abundant divalent cation in living cells, with a total cellular concentration between 14 and 20 mM (Schmitz et al., 2007). The substrate curve demonstrated that the apparent KM for adenosine was around 1.13 ± 0.07 mM and the estimated Vmax for adenosine deamination was 2.61 ± 0.054 NH3 min−1 mg−1 protein in T. vaginalis. The kinetic data obtained in this study are in accordance with other studies related to ADA activity, although there are some variations of KM among different ADA members. The KM value of H. dromedarii ADA2 was estimated to 0.5 mM adenosine (Mohamed, 2006), which is relatively close to several ADAs from different sources, such as rat brain (0.45 mM) (Centelles et al., 1988), bovine brain (0.4 mM) (Lupidi et al., 1992), human (0.46 mM) and chicken liver (0.33 mM) (Iwaki-Egawa & Watanabe, 2002). However, lower KM values were reported for ADA activity from mice intestine (0.023 mM) (Singh & Sharma, 2000) and from the sand fly Lutzomyia longipalpis (0.01 mM) (Charlab et al., 2000). Additional data on biochemical characterization revealed the strong preference of the T.

There is still no report on the purification of full-length squal

There is still no report on the purification of full-length squalene synthase by Ni-NTA chromatography, and so the recombinant protein was purified by Q-Sepharose followed by phenyl superose

and was analysed by 10% SDS-PAGE (Fig. 3a). The recovery of the enzyme activity in the various steps of selleckchem its purification procedure is presented in Table 1. Uninduced culture and BL21 (DE3) E. coli cells transformed with the pET-28(a) vector without the SSN gene were used as control. The specificity of the protein was further validated by Western blot by probing it with His antibodies because the expressed protein has a His6-tag attached to its C-terminal. The His antibodies specifically binds with His-expressed protein in the total cell lysate, pellet, supernatant and partially purified samples, while no cross-reactivity was detected in negative controls, which confirms the expression of His-tag protein in samples (Fig. 3b). To demonstrate that the overexpressed recombinant LdSSN protein actually has SSN activity, the conventional radioactive assay was performed using purified recombinant LdSSN protein. The pH dependence, thermal

stability and effect of denaturants (urea and GdmCl) were studied on recombinant LdSSN protein. Similar to most other SSNs, LdSSN showed activity selleck chemical in alkaline pH (Belingheri et al., 1991; Shechter et al., 1992). The pH optimum for the LdSSN was observed as 7.4, which was in comparison with trypanosomal, rat and daffodil SSN (Belingheri et al., 1991; Shechter et al., 1992; Sealey-Cardona et al., 2007), but was slightly

higher than the value of 7.2 reported for the yeast enzyme (Zhang et al., 1993). Moreover, a plateau was observed in the region of pH 7–7.8. The enzyme retained >80% activity in the buffer range of MOPS NaOH (Fig. 4a). Thermal stability of SSN varies in different organisms. The temperature optimum may be as high as 60 °C in Thermosynechococcus elongatus BP-1 (Lee & Poulter, 2008) and as low as 37 °C in other organisms. LdSSN showed maximum activity at 37 °C, whereas it exhibited Fenbendazole 83% and 88% activity at a temperature of 30 and 45 °C. LdSSN was found to be temperature sensitive as compared with other SSN, as it loses 85% of its activity at 60 °C (Fig. 4b). The effect of denaturants (urea and GdmCl) on LdSSN was assessed by incubating the enzyme at different concentrations of denaturants. The enzyme lost 81% and 86% of its activity at a concentration of 2 M urea and 0.3 M GdmCl, respectively (Fig. 4c and d). The enzyme loses >50% activity at a concentration of 1 M urea and 0.2 M GdmCl. The activity of the protein is more sensitive towards GdmCl than that of urea; this might be due to the ability of GdmCl to disturb the electrostatic interactions. The loss of activity can be due to unfolding of the enzyme, or due to disruption of the active-site microenvironment in the presence of denaturant molecules or due to preferential binding of molecules on the surface of LdSSN.

Moreover, unbiased microarray expression analysis showed that Cxc

Moreover, unbiased microarray expression analysis showed that Cxcl10 was among 112 transcripts in the neocortex upregulated at least threefold in both TBI and ageing TgSwe

mice, many of them involved in inflammation. The identity of the Cxcl10+ cells remains unclear but flow cytometry showed increased numbers of activated microglia/macrophages as well as myeloid dendritic cells in the TBI and experimental autoimmune encephalomyelitis models. It is concluded that the Cxcl10+ cells appear in the inflamed central nervous system and may represent a novel population of cells that it may be possible to target pharmacologically in a broad range of neurodegenerative conditions. “
“We combined computational modeling and experimental measurements to determine the influence of dendritic structure on the diffusion of Docetaxel intracellular chemical signals in mouse cerebellar Purkinje cells and hippocamal CA1 pyramidal cells. Modeling predicts

that molecular trapping by dendritic spines causes diffusion along spiny dendrites to be anomalous and that the value of the anomalous exponent (dw) is proportional to spine density in both cell types. To test these predictions we combined the local photorelease of an inert dye, rhodamine dextran, with two-photon fluorescence Baf-A1 imaging to track diffusion along dendrites. Our results show that anomalous diffusion is present in spiny dendrites of both cell types. Further, the anomalous exponent is linearly related to the density of spines in pyramidal cells and dw in Purkinje cells is consistent with such a relationship. We conclude that anomalous diffusion occurs in the dendrites of multiple types of neurons. Because spine density is dynamic and depends on neuronal activity, the degree of anomalous diffusion induced by spines can dynamically regulate

the movement of molecules along dendrites. “
“Preconditioning rat hippocampal–entorhinocortical (HEC) slice or cerebellar cell cultures with moderate concentrations of ethanol (20–30 mm) neuroprotects against pro-inflammatory proteins such as HIV-1 glycoprotein 120 (gp120) or amyloid-β. The neuroprotective mechanism of ethanol is unclear, but it conceivably involves sensorstransducerseffectors, analogous Urease to other preconditioning modalities. We initially found that the preconditioning augmented two likely heat shock protein (HSP) ‘effectors’, HSP70 and HSP27, and that precluding HSP upregulation abolished neuroprotection. Here we examined whether pro-survival kinases are transducers potentially leading to HSP effectors. In cerebellar cultures, protein kinase C (PKC) activity increased modestly after 2 days of 30 mm ethanol and was significantly induced after 6 days, when neuroprotection against gp120 becomes manifest.

It has advantages over restriction endonuclease–based methods and

It has advantages over restriction endonuclease–based methods and is usually rapid.

Typically, recombineering uses long PCR primers (c. 65 bases), each of which contains a small region of target homology (c. 45 bases). We have developed a simple, albeit somewhat less rapid, strategy to create recombineering substrates that can use primers of ≤ 35 bases for all steps. The regions of homology can be several hundred base pairs in length to (1) increase the chance of obtaining the desired clone and/or (2) allow coliphage-based recombineering in some non-Escherichia coli bacteria. The method uses cloning techniques to construct a template for the generation of the recombineering substrate. Because the template is made from cloned DNA segments, the segments (including those for the homology regions) can be readily changed. During construction of the template plasmid, potential background http://www.selleckchem.com/products/pexidartinib-plx3397.html transformants arising from the vector SB431542 without insert are significantly reduced by cloning each segment with two restriction endonucleases that produce noncompatible ends. We have used this method to change the bla gene of pACYC177 to aadA, to add the MCS-lacZα region from pBBR1MCS to IncQ plasmid vectors, and to make an oriTIncP-aacC1 cassette and add

it to a plasmid. Recombineering (genetic engineering by homologous recombination using the red genes of bacteriophage λ or the recET genes of the defective Rac prophage) is a powerful and convenient method for changing or cloning DNA (Murphy, 1998; Zhang et al., 1998; Datsenko & Wanner, 2000;

Yu et al., 2000; for reviews, see Muyrers et al., 2001 and Court et al., 2002). Unlike the commonly used molecular cloning techniques that are based on ligation of DNA in vitro, the covalent linkage of genetic determinants in recombineering occurs in vivo by homologous recombination. Because recombineering can use any nucleotide sequence as a target, it is not limited by the chance occurrence of restriction endonuclease cleavage sites in the target DNA. Homologous recombination by the λ red or the recET recombination systems is independent of see more the recA gene of the host (Muyrers et al., 2001; Court et al., 2002). Only c. 45 bp of homology is required for homologous recombination. The proteins encoded by the λ red system (Exo, Beta, and Gam) will be highlighted here. A major function of Gam is to inhibit the degradation of linear DNA molecules by the Escherichia coli RecBCD exonuclease (Murphy, 1991) and the SbcCD exonuclease (Kulkarni & Stahl, 1989). Exo (an exonuclease) processes linear duplex DNA molecules so that they can participate in red-mediated homologous recombination (Little, 1967; Carter & Radding, 1971). Beta helps catalyze homologous recombination by binding to the exposed single strands of the processed linear DNA molecules and promoting the annealing of complementary strands (Carter & Radding, 1971; Muniyappa & Radding, 1986).

One block consisted of ten 32-s

trials The middle third

One block consisted of ten 32-s

trials. The middle third of each tracking pattern was repeated and identical across practice and retention (Boyd & Linsdell, 2009). The pattern was unknown to the participants and was constructed from the polynomial equation described by Wulf & Schmidt (1997) with the following general form: The middle (repeated) segment was constructed by using the same coefficients for every trial (b0 = 2.0, a1=−4.0, b1 = 3.0, b2=−3.6, a3 = 3.9, b3 = 4.5, a4 = 0.0, b4 = 1.0, a5=−3.8, b5=−0.5, a6 = 1.0 and b6 = 2.5). The first and third segments of the tracking pattern were generated randomly using coefficients ranging from −5.0 to 5.0. A different random sequence was used for both the first and third segments of every trial. There were 10 separate reversals in the direction CYC202 cell line in each third of the tracking pattern. The random and repeated patterns were equated for difficulty by ensuring that the overall excursion of each random sequence fell within a range of that required by the repeated

sequence. Neither the trajectories of the target nor the participants’ movements left a visible train on the screen and thus participants could not visualize the entire pattern. The same sets of trials were practised by all of the participants to ensure uniformity so that the random segments were the same for each participant. Participants were not informed of the repeating sequence but were instructed daily to track the target as accurately as possible by controlling the position of the cursor with the joystick. Transcranial magnetic stimulation was delivered find more with a Magstim Super Rapid2 stimulator using a 70-mm figure-of-eight air-cooled coil (Magstim Company Ltd., Whitland, UK). Participants were seated in a semi-reclined dental chair with their arms bent and supported by armrests. The TMS coil was orientated tangentially to the scalp with the handle at 45° to the midline in a posterior lateral orientation. Prior to the experiment, high-resolution anatomical magnetic resonance images (MRIs) were acquired for each participant (TR = 12.4 ms, TE = 5.4 ms, flip

angle θ = 35°, FOV = 256 mm, 170 slices, 1-mm thickness) at the UBC MRI Research Centre on a Philips Achieva 3.0 T whole body MRI scanner (Phillips Healthcare, Andover, MD, USA) using a sensitivity encoding head coil (SENSE). These images were then imported into the BrainSight™ TMS neuronavigation software (-)-p-Bromotetramisole Oxalate (BrainSight 2.0, Rogue Research Inc., Montreal, QC, Canada) to allow for stereotaxic registration of the TMS coil with the participants’ anatomy for online control of coil positioning during each session and across days. Surface electromyography over the participants’ right flexor carpi radialis (FCR) was monitored using the evoked potential unit of the Super Rapid2 control unit (Magstim Company, Ltd) (Boyd & Linsdell, 2009). Initially, the FCR representation was marked on the participants’ anatomical MRI as the medial edge of the left ‘hand knob’.