Several studies compare LDR BT as

Several studies compare LDR BT as SCH772984 order standard treatment vs. HDR BT, with some contradictive results. A recent meta-analysis pooled the results of randomized studies and concludes no significant differences for survival and LC (19). In interpreting these results, it is necessary to keep in mind the range of radiation and BT technologies

used in these studies. PDR seems to be a good compromise between LDR and HDR with radiobiologic advantages of LDR and technical advantages of HDR. Only one prospective study has compared continuous LDR BT and PDR BT for cervical carcinoma: 166 patients were analyzed prospectively, 57 in the PDR BT arm. The dose rate was similar in both groups (66 cGy/h in LDR and 70 cGy/h in PDR arm). No differences were found for severe late toxicity. The actuarial 3-year OS rate was 75% for both groups, with no significant differences in 3-year DFS for the PDR BT group (70% vs. 57%, p = 0.19) (20). Only one randomized prospective study suggests the impact of LDR variations (21), with 204 patients with Stage I and limited Stage II cervical cancer randomized to receive one of two preoperative BT LDRs (0.4 and 0.8 Gy/h). The investigators reported a greater late complications rate with

higher dose rate (38 cGy/h vs. 73 cGy/h), with no impact on survival. Our results do not support this finding as we find a low complication rate with a median dose rate of 65 cGy/h: 2.6% of gastrointestinal tract complications, this website 4.4% urinary tract severe toxicity, and 1.3% complete obliteration of the vagina. These toxicity rates were in accordance with those established with LDR. In the review by Barillot et al. (22), 4% of late severe urinary toxicity and 2–4% of gastrointestinal oxyclozanide tractus (35% for locally advanced cancer) were established. We acknowledge the limitations of this study owing to its retrospective assessment of toxicity; however,

with 50.6% Grades 1 and 2 late vaginal effects, our rate is less than that reported by Potter et al. (23) in a large series of HDR BT (78%). In the multivariate analyses on outcomes, classical clinical factors such as negative nodal involvement are correlated with the 5-year LC but also the use of 3D-based planning BT. Interest in BT 3D imaging planning has increased and represents currently one of the most important developments in gynecologic BT. Recently, guidelines have been published by the GEC ESTRO [14] and [24]. However, up until now, limited clinical evidence has been published demonstrating the impact of 3D BT. Chargari et al. (25) have been the first to report their experience with MRI-based intracavitary PDR BT so far, for 45 patients with locally advanced cervical carcinoma. The 2-year OS was 78% without any Grade 4 toxicity and with only one Grade 3 toxicity with a vesicovaginal fistula.

Although studies with KO mice often suffer from some weaknesses 4

Although studies with KO mice often suffer from some weaknesses 42 and 43••], they have undoubtedly contributed

enormously Seliciclib manufacturer to our understanding of how genes influence behavior. It should be noted, however, that these studies do not necessarily shed light on the question what makes individuals different from each other, simply because natural populations are not necessarily polymorphic for the genes that have been studied in KO mice [44]. Fortunately, new tools have become available or are currently being developed that aid or will aid enormously in the task of identifying genes responsible for individual differences. The Collaborative Cross, which aims to develop hundreds of recombinant inbred strains, is one example [45]. The Diversity Outbred mouse population is another one [46]. The extended family of BXD recombinant inbred strains [47] is already being used in many studies. In general, therefore, we are seeing exciting developments in the wider

field of behavior genetics and the future appears bright. Some dark clouds remain, however: In my considered opinion, defining phenotypes is currently the most important and most pressing problem, both for animal behavior genetics and psychiatric genetics. Ever since the PI3K signaling pathway landmark study of Crabbe et al. was published in 1999 [48••], researchers have worried about the replicability of behavioral data obtained with genetically defined animals in standardized tests. Crabbe and colleagues tested a number of inbred strains, as well as one KO mutant, simultaneously in three different laboratories on a battery of carefully standardized behavioral tests. The results came as a shock to many in the field: large differences were found between the results obtained in the different laboratories

for some of the tests. The most striking result involved anxiety measured on a plus maze, where large inter-laboratory differences cropped up. While these data give pause for thought, it would appear that the initial reaction to them was too extreme. Crabbe et al. did most emphatically not show that behavioral research with mice is not replicable. Hydroxychloroquine price In fact, the more surprising result of their study was that so many behaviors replicated very well [49]. As they observed a few years later, ‘Only on a test of anxiety was the variation among labs close to the magnitude of genetic variation’ [50]. In later studies, the same authors also showed that behavioral test results obtained with standardized inbred strains are stable, not just between different laboratories, but even over decades [51••]. In short, the problem with behavioral phenotypes is not the replicability of results, because with adequate care and standardization (apparently even including the sex of the experimenter [52]), this can be achieved.

As a substantial number of the substances have already been teste

As a substantial number of the substances have already been tested for some of the methods, it is expected that the remaining data gaps will be filled soon. This will allow re-assessment of already proposed testing strategies, e.g. by Bauch et al., 2012, Gomes et al., 2012, Nukada et al., 2012, Natsch et al., 2013 and Jaworska et al., 2013, with new data. Once the data for the eight test methods will be available, a testing strategy will be composed addressing the specific purposes and needs as described above. Driven Cyclopamine by the mechanistic understanding and supported by data analysis specialists, data mining and other statistical tools will be used to combine test method data in an objective and transparent

way to selleck chemical obtain a predictive testing strategy that will be made publicly available. Predictive performance will be assessed correlatively/probabilistically against the reference human and LLNA data or a combination thereof. Although efficiency and other factors, such as availability or duration, may – at least at this stage – not be accounted for, it is nevertheless expected

that the strategy will comprise a limited number of test methods. In the third phase of the framework, applicability domain issues specifically relevant for substance used by cosmetic industry will be addressed. It is anticipated that for inherently problematic substance types, such as natural extracts, dyes or polymers, further data may be required Protein kinase N1 in order to provide sufficient evidence that the testing strategy works for these substance types or to optimise the adaptation of the strategy. The resulting non-animal testing strategy for skin sensitisation potency predictions will be combined with bioavailability and skin metabolism data, exposure consideration and in exceptional cases with data from T cell activation assays, to satisfy the ultimate goal of a data integration approach for skin sensitisation safety assessment of cosmetic ingredients. The authors declare that there are no conflicts of interest. Transparency document.

We would like to thank Pierre Aeby for his support of the method evaluation leading to the workshop. Furthermore, we would like to thank Silvia Casati (EC, EURL-ECVAM) for her active participation in the workshop. “
“Cutaneous malignant melanoma (CMM) presents significant morbidity and a high mortality rate and is the most dangerous of all common skin cancers because of its propensity to metastasize. The number of melanoma cases worldwide is increasing faster than any other cancer. In contrast to other types of cancer, CMM frequently affects young individuals, with a mean age of 50 years (Ferrari et al., 2008). The treatment of patients with advanced melanoma remains an important issue to be investigated. The same chemotherapies that are effective in numerous types of cancer are largely ineffective in melanoma (Huncharek et al., 2001).

In contrast to baseflow conditions,

stormflow waters refl

In contrast to baseflow conditions,

stormflow waters reflect the acidic nature of precipitation in the region, including NO3 concentrations derived largely from sources outside the watershed (Fig. 4) and the slightly enhanced solubility of trivalent metals such as Al and the REEs. The concentration of SO4 is less variable between events and likely controlled by the oxidation of common sulfide-rich minerals such as pyrite. As noted above, the Raymondville sampling site (RY on Fig. 1) is intriguing because of its anomalous geochemistry compared with AG-014699 molecular weight other sampling sites during storm flow (Fig. 3 and Fig. 4). This was particularly evident during the stormflow after Tropical Storm Irene, but not apparent during the baseflow sampling event. In particular, the Raymondville sampling site stands out during the stormflow sampling as the only site to have an alkaline pH (8.21), the largest concentrations of the anions CO3 and SO4, and the largest concentrations Cetuximab in vitro of Ba, Ca, Cl, K, Mg, Na, Rb, Si, and Sr (∼3 times baseflow concentrations; Fig. 3 and Fig.

4). Slight decreases in the trivalent cations were also found in the Raymondville stormflow sample when compared to samples collected up- and downriver. These chemical trends have been duplicated in another study which sampled Raquette River waters at Raymondville weekly for an entire year (Laboso et al., 2014), indicating control by a continuing, but sporadic, process. Review of land use south of the Raymondville sampling site on the Raquette River indicates that a large quarry (∼1.3 km × 0.4 km) exists 6 km upriver at Norfolk

(Fig. 6). The quarry is located on east bank of the Raquette River and produces a variety of crushed stone products for construction and other purposes. The rock quarried here is the Ordovician Ogdensburg Dolostone. Previous studies have indicated that evaporitic horizons exist in the dolostone and samples from water wells in nearby Louisville and on the Akwesasne Mohawk Nation east of Massena which penetrate Histone demethylase it, have an enrichment in soluble elements such as B, Ca, Br, K, Li, Rb, and, particularly, Sr (Chiarenzelli et al., 2007 and O’Connor et al., 2010). A view of the quarry from Google Earth on May 26, 2011 (Fig. 6) indicates that it has standing water in low areas and stockpiles of a variety of crushed stone products. In addition, a plume of material, presumably fine rock powder, can be seen entering the river there and is carried downstream. On that day at the Massena airport 0.23 in. of rain fell. The monthly total at that point was 3.96 in. compared to a long-term average of 2.56 in.

Consequently, the interrelations of representative statistical pe

Consequently, the interrelations of representative statistical periods also change. Instead of the standard relations Tmax ≈ T1/10 ≈ Ts ≈ 1.1 − 1.2 Tm, these relations behind the breakwater tend to Tmax ≈ T1/10 ≈ Ts ≈ 1.5 Tm. It was also concluded that the mean wave periods calculated by

both the statistical approach (zero up-crossing) Tm and the spectral approach T0.2 have approximately Rucaparib research buy the same values behind the breakwater, i.e. wave spectral deformation does not affect the calculation of the mean spectral period T0.2. The mean spectral period T0.2 depends on the relative submersion Rc/L0.2 − i and is reduced as submersion approaches zero for both submerged and emerged breakwaters. It is estimated that the greatest reduction in period T0.2 when waves cross

the smooth breakwater occurs when the relative submersion is Rc/L0.2 − i ∼ 0 and amounts to ∼ 70% of the value of the incoming mean period. The peak period Tp increases or remains the same when the waves cross the smooth submerged breakwater. As far as the emerged breakwater is concerned, there is a dependence of the peak period Tp, and the relative submersion Rc/L0.2 − i. By increasing the relative submersion, the peak period Tp increases by up to 35% in relation to the incoming peak period. The empirical model is formed for estimating the reduction in the mean spectral period when the waves cross submerged and emerged smooth breakwaters. For the incoming wave parameters and the depth in the breakwater crown, the model provides the values for the reduction coefficients K0.2R−TKR−T0.2. As the model was derived from a restricted Cepharanthine number of measurements, additional

measurements will be necessary, particularly in the zone of relative submersion Rc/L0.2 − i ∼ 0. Measured reduction coefficients of the mean period agree well with the calculated values. List of symbols: Hmax maximum wave height, [m], (zero up-crossing), “
“The first data on the Barents Sea sipunculan fauna were reported by N. K. Zenger in 1870 (Zenger 1870). In the first half of the 20th century, two reviews of the Gephyrea of USSR seas were written (Vagin, 1937 and Zatsepin, 1948). At that time, the term Gephyrea was used for the group of marine coelomic worms without obvious segmentation – sipunculans, priapulids and echiurids. Extensive data on the Barents Sea Sipuncula is given in the monograph by G. V. Murina (1977) on the sipunculan fauna of Eurasian Arctic and boreal waters. Sipuncula is a relatively species-poor phylum consisting of about 150 species and subspecies worldwide (Cutler 1994); the checklist for Arctic seas has fewer species. According to these publications there are 7 Sipuncula species living in the central part of the Barents Sea and East Murman inshore waters: Phascolion strombus strombus (Montagu 1804), Golfingia elongata (Keferstein 1863), G. margaritacea margaritacea (Sars 1851), Nephasoma eremita (Sars 1851), N. improvisa (Théel 1905), N.

Immunoreactive bands were visualized using a chemiluminescence re

Immunoreactive bands were visualized using a chemiluminescence reagent (Amersham Biosciences, Buckinghamshire, UK), followed by autoradiography. β-actin was used as the loading control. Densitometry of various analyte proteins and their respective loading controls from the same blot was performed using Image J 1.43 (NIH) software. Relative optical density was calculated by

dividing the densitometry of analyte(s) protein with the respective loading control. The levels of BPDE-DNA adducts were detected by immunohistochemical staining for BPDE-DNA adducts in formalin-fixed, paraffin embedded 5 μm tissue sections as described previously [14]. Sections were incubated with anti-BPDE antibody (1:30

dilution). Detection was done using Vectastain ABC kit (Vector Laboratories, Burlingame, CA, USA). Diaminobenzidine Inhibitor Library cost (DAB) was employed as the chromogenic substrate, and slides were counterstained with Mayer’s haematoxylin. Images were captured with Zeiss Microscope (Imager Z1) to which an Axiocam MRc5 digital AG-014699 solubility dmso camera was attached. Quantitative analysis of the images (magnification X 400) was performed by IHC profiler [15], which is an open source plug-in for the quantitative evaluation and automated scoring of immunohistochemistry images of tissue samples. This modified digital image analysis is based on protocols adopted earlier [16]. IHC photomicrographs were used for developing semi-automated analysis protocol, namely IHC profiler [15]. As a first step, a color de-convolution plug-in was used to un-mix the pure DAB and haematoxylin stained areas that left a complimentary image. The pixel intensities of separated DAB images range from 0 to 255. Value 0 represents the darkest shade whereas 255 represents the lightest shade of the DAB brown color in the image. To select the DAB-stained (brown) nuclei, the threshold feature of the Image J 1.43 (NIH) software was used. Further to assign an automated percentage of pure DAB staining patterns in the nucleus, a macro was developed and plugged in the Image J 1.43 (NIH) software to obtain an automated counting

of the pixel wise percentage contribution of high, medium and low positive pixels/intensity in an image i.e. the number of pixels of a specific intensity value Amrubicin vs. their respective intensity zone. For measurement of BPDE-DNA adducts [similar areas of tissue sections (mm2) and number of cells (∼800 cells/section/animal)], total intensity (%) [of nuclei containing percentage of high, medium and low intensity] was analyzed within different treatment groups. However, apoptosis was measured in terms of total apoptotic nuclei intensity as well as percentage of apoptotic positive and negative cells in similar areas of tissue sections (mm2) and number of cells (∼800 cells/section/animal) in different treatment groups.

In this review we highlighted

two potential innate inflam

In this review we highlighted

two potential innate inflammatory mechanisms that may lead to development of synovitis in OA, the TLR pathway and the complement cascade (Fig. 3). Furthermore, we highlighted the roles of cytokines AC220 mouse and chemokines that play a role in the initiation and perpetuation of synovitis and OA symptoms. These pathways and mediators also may impact cartilage matrix homeostasis and peri-articular bone remodeling. In addition, the products associated with synovial inflammation may serve as surrogate markers of disease activity or responses to therapeutic interventions. Further understanding of mechanisms promoting synovial inflammation in OA may lead to identification of novel therapeutic targets for controlling symptoms and slowing structural progression in this disabling

joint disease. This study was supported by 1K08 AR057859-02, Mentored Clinical Scientist Career Development Award, from the National Institute of Arthritis, buy PTC124 Musculoskeletal and Skin Diseases (CRS). “
“Linear bone growth involves the replacement of a cartilaginous template by mineralized bone through endochondral ossification. This growth process is orchestrated by various actions at the growth plate, a developmental region consisting of chondrocytes in distinct cellular zones. The proliferation, hypertrophy and apoptosis of these growth plate chondrocytes are regulated by a tight array of factors ensuring effective cartilage JAK inhibitor mineralization and thus longitudinal growth [1]. Hydroxyapatite (HA) crystals form associated with the trilaminar membrane bound matrix vesicles (MV) which in the growth plate are localised to the mineralized longitudinal septae and form from the plasma membrane of the terminal hypertrophic chondrocytes [2]. Mineralization is a biphasic process which is under tight control so as to

ensure levels of calcium (Ca2 +) and inorganic phosphate (Pi) are permissive for effective HA formation [2]. Three molecules have been identified as imperative in controlling levels of the mineralization inhibitors inorganic pyrophosphate (PPi), and osteopontin [2] and [3]. These are alkaline phosphatase (ALP), a nucleotide pyrophosphatase/phosphodiesterase isozyme (NPP1), and the Ankylosis protein (ANK). However, mechanisms beyond the supply and hydrolysis of PPi likely exist to control chondrocyte matrix mineralization. Once such mechanism could involve matrix extracellular phosphoglycoprotein (MEPE, OF45). This was originally isolated and cloned from tumors of oncogenic hypophosphatemic osteomalacia (OHO) as a candidate substrate for phosphate-regulating gene with homologies to endopeptidases on the X chromosome (PHEX) [4]. MEPE is a 56–58 kDa SIBLING (small integrin-binding ligand N-linked glycosylated) protein along with dentin matrix protein 1 (DMP1), osteopontin (OPN), dentin sialophosphoprotein (DSPP) and bone sialoprotein (BSP) [5].

Subject-specific movement parameters were also included as regres

Subject-specific movement parameters were also included as regressors of no interest. Participant-specific parameter estimates (β values) were calculated at each voxel across the brain. The parameter estimates were then entered into a second level random effects analysis, where one-sample t-tests were applied to every voxel. Initial statistical thresholding was applied using a threshold of p < .001, uncorrected for multiple comparisons. Activations were considered to be statistically significant only if they survived FWE correction at either the peak or cluster level. For a priori anatomical ROIs, FWE correction was applied using small volume

correction ( Frackowiak et al., 2004) within pre-defined anatomical masks (see Section 2.7). Roxadustat concentration Although not our primary interest, given the results of Park et al. (2007), we also looked for adaptation effects. Here we contrasted trials where the two scenes were perceived to be the same with those that were perceived to be different (including both closer and further away), despite the stimuli being physically identical during any one trial. The trials were divided into these two conditions for modelling both the first and second scene presentations. In all other respects, the experimental design was identical to that described above. We first used a

whole-brain analysis to localise regions which displayed an overall adaptation effect between the first and second scene presentations, regardless of condition. We then conducted more in-depth Molecular motor adaptation analyses using ROIs, as described below. The statistical thresholds were identical to those described above. Given the (limited) previous literature on the functional neuroanatomy of BE, our a priori hypothesis was that the HC would be involved in the BE effect, and that the PHC and RSC might also be implicated. Each of these ROIs was manually defined on the normalised group average T1-weighted structural MR image using the Duvernoy anatomical atlases for guidance ( Duvernoy, 1999, 2005). These anatomical ROIs were used in MarsBar ( analyses, where a finite impulse response

(FIR) model ( Dale, 1999; Ollinger et al., 2001) was fitted to the data in order to probe the time-course of responses in ROIs. Four time-windows of 2 sec each were modelled, time-locked to the onset of the first scene presentation. These ROIs were also used for small volume correction within SPM. Based on the whole-brain adaptation analysis described above, we determined that early visual cortex (VC) was a target for further ROI-based analyses. We therefore established a VC ROI using a contrast that was orthogonal to the adaptation analysis (i.e., all scenes presented on the first trial only compared to the implicit baseline). This ROI was used for further adaptation analyses, and for the HC–VC dynamic causal modelling (DCM) analysis described below.

Protein was extracted from fresh

Protein was extracted from fresh PF-02341066 in vitro tobacco leaves by homogenization in extraction buffer (200 mmol L− 1 Tris–HCl (pH 8.0), 100 mmol L− 1 NaCl, 400 mmol L− 1 sucrose, 14 mmol L− 1 isoamyl alcohol, 1 mmol L− 1 phenylmethylsulfonyl fluoride (PMSF) and 0.05% Tween-20). The extract was centrifuged at 12,500 r min− 1 for 20 min at 4 °C. The protein concentration of the supernatant was determined using the Bio-Rad protein assay. The protein samples were mixed with

50 μL of 3 × sodium dodecyl sulfate (SDS) loading buffer (Bio-Rad) and boiled for 10 min, and 8 μL of each sample was subjected to SDS-polyacrylamide gel electrophoresis (PAGE) on 12% Tris–glycine gels (Invitrogen). Protein bands were transferred to a Poly vinylidene fluoride (PVDF) membrane. After blocking with 5% BSA for 1 h at room temperature, the

blots were incubated overnight at 4 °C with antiserum (1:10,000 dilution) in the presence of 1% BSA, washed three times (15 min each), and incubated with 1:30,000-diluted alkaline phosphate-conjugated anti-rabbit IgG for 1 h at room temperature. The reaction was visualized with a BCIP/NBT color development substrate (Promega, Inc.). The anti sera used were raised in rabbits. Two methods Everolimus in vivo were used to analyze glyphosate tolerance in transgenic tobacco plants. For the leaf spraying experiment, 6 to 8-leaf-stage transgenic plants grown in the green house were sprayed with the herbicide Roundup (isopropylamine salt of glyphosate as active ingredient), 41.0% (w/v) at doses of 0.8–1.0 L ha− 1. T1 progeny seeds of transgenic tobacco containing gat, G2-aroA, or gat/G2-aroA were germinated on MS medium supplemented with 0, 0.2, 1.0, 5.0, and 10.0 mmol L− 1 glyphosate. Seedlings were grown in growth chambers at 25 °C with 60%–70% relative humidity and a photosynthetic photon flux density of 24 μmol m− 2 s− 1 with a 10-h photoperiod. The growth status

and viability of transgenic plants were evaluated after culturing for 4 weeks. The gat gene was amplified by PCR using corresponding primers and template. After sequencing confirmation, the gene was inserted into pG2 to form plant expression vector p2301G2-GAT. In this vector, gat and G2-aroA genes were driven in tandem by a CaMV35S promoter Molecular motor with two enhancers and terminated with a NOS terminator at their 3′ ends. The T-regions in p2301G2-GAT also harbored 35SP::nptII::35SpolyA to provide kanamycin resistance. The structure of p2301G2-GAT is shown in Fig. 1. A total of 52 independent transgenic tobacco (N. tabacum cv. NC89) lines were generated by Agrobacterium-mediated gene transformation. The transgenic plants with G2-aroA and gat were named G2-GAT. Southern blotting, RT-PCR, and Western blotting analysis showed that the specific bands were present in tested samples ( Fig. 2, Fig. 3 and Fig.

Any trials on which

Any trials on which selleck chemicals llc a participant provided this response were discarded from the subsequent analysis, as were trials on which participant failed to provide a response to either of the ratings [mean number of excluded trials 1.53 (SD 2.5)]. Participants then had 2 sec to rest before the start of the next trial. Following the scoring procedure of Intraub and Richardson (1989), each response

was scored from −2 to 2 where −2 meant “much closer-up”, −1 meant “a little closer-up”, 0 meant “the same”, 1 meant “a little further away”, and 2 meant “much further away”. The mean score across all trials was calculated for each participant, providing an overall BE score. This score indicates the degree of bias towards one

response over another. If participants show no bias in response, the score will be 0. However, if they display a BE effect, the score will be negative, due to the greater number of closer responses. In order to determine whether the group of participants as a whole displayed a significant BE effect, we compared the BE scores to 0 C59 wnt molecular weight using a t-test. We also performed a second analysis where we investigated the proportion of each response type (Closer, Same, Further), ignoring the degree of subjective distance (i.e., whether it was “much” or “a little” further/closer). For this analysis we calculated the percentage of response trials falling into each of the three categories for each participant, and compared them using a one-way analysis of variance (ANOVA). MRI data were Depsipeptide chemical structure collected

using a 3 T Magnetom Allegra head-only MRI scanner (Siemens Healthcare, Erlangen, Germany) operated with the standard transmit-receive head coil. Functional MRI data were acquired in one session with a BOLD-sensitive T2*-weighted single-shot echo-planar imaging sequence which was optimised to minimise signal dropout in the medial temporal lobe (MTL) (Weiskopf et al., 2006). The sequence used a descending slice acquisition order with a slice thickness of 2 mm, an interslice gap of 1 mm, and an in-plane resolution of 3 × 3 mm. Forty eight slices were collected covering the entire brain, resulting in a repetition time of 2.88 sec. The echo time was 30 msec and the flip angle 90°. All data were acquired at a −45° angle to the anterior–posterior axis. In addition, field maps were collected for subsequent distortion correction (Weiskopf et al., 2006). These were acquired with a double-echo gradient echo field map sequence (TE = 10 and 12.46 msec, TR = 1020 msec, matrix size 64 × 64, with 64 slices, voxel size = 3 mm3) covering the whole head. After these functional scans, a 3D MDEFT T1-weighted structural scan was acquired for each participant with 1 mm isotropic resolution (Deichmann et al., 2004). Neuroimaging data were analysed using SPM8.