Cerebellar development involves a spurt of proliferation in external granule layer (EGL) in response to shh, causing granule neuron precursors (GNPs) to proliferate. These cells subsequently differentiate into granule neurons in the inner granule layer (IGL). F3, a CNTN family molecule, can interact with NrCAM to switch GNPs from proliferation to differentiation. We aim to identify the role of NrCAM in the sonic hedgehog response in GNPs.GNPs were extracted from wildtype and NrCAM mutant P5 cerebella using Percoll gradient centrifugation. Proliferation response to shh was measured using EdU in presence/absence of F3-Fc. GNPs treated with shh/SAG were stained with antibodies against Arl13b and smo to look for differences in cilia size and smo occupancy after different treatment times.-/- and wildtype GNPs both proliferated equally in response to shh. F3 was found to block the proliferation response in wildtype but not in NrCAM-/- GNPs. F3 also failed to affect proliferation in SmoA1 GNPs with a constitutively active smo suggesting that the F3-NrCAM mediated block lay upstream of Smo. NrCAM was detected in wildtype cilia and Smo localization was affected in NrCAM-/- GNPs. No differences in cilia length were observed.NrCAMOur results suggest that NrCAM affects shh-mediated proliferation by controlling smo movement into the cilium.


Recent advances in our understanding of the contributions of gut microbiota to host metabolism have renewed research interest in how distortions in microbiota activities correlate with health status. While extensive microbiota surveys have already revealed the complexity of microbiota dynamics there is a need to better understand mechanisms through which microbiota can affect host metabolism. We have recently shown that the amount of energy available to the host after intake of resistant maltodextrin varies among individuals and is associated with microbiota. Here we hypothesized that microbiota i) is associated with dietary habits, ii) correlates with colorectal cancer (CRC) risk and iii) can affect the methylation status in nearby gut epithelium, which might be a mechanism contributing to colorectal carcinogenesis.In 126 human volunteers undergoing a screening colonoscopy we determined associations between dietary intake, gut microbiota composition, epithelial methylation status and polyp prevalence. We determined: i) dietary habits using 3-day food records, FFQ (Block 98), and Meat Module Questionnaire; ii) medical history , iii) microbiota composition in a fecal sample collected before and multiple biopsy samples collected during the colonoscopy, during which we determined iv) polyp status. Microbiota composition was analyzed in DNA extracted from stool and biopsy samples using a 16S rRNA based approach. Sequence reads were binned into operational taxonomic units (OTUs) using ESPRIT and analyzed using QIIIME. We then performed a discriminant analysis to identify a microbiota pattern associated with polyp prevalence. For a subset of fecal samples a shotgun metagenomic analysis was undertaken. An exploratory methylation analysis was performed in DNA extracted from 12 biopsy samples using the Infinium HumanMethylation450 beadchip.Various microbiota associations with dietary intake as well as differences in the presence of OTUs between subjects with polyps and controls (p<0.01) were observed, with the most significant of these differences detected in subjects with high risk polyps. A predictive model based on 27 OTUs was effective in identifying subjects with at least one polyp (AUC =0.81). Butyrate producing bacteria appeared decreased among polyp cases. Methylation status at multiple sites was associated with polyp status, which correlated with specific microbiota differences.Our observations confirm microbiota contributions to host metabolism. Larger studies in targeted populations will be needed to determine more details of the complex mechanisms of interactions between microbiota activities and host metabolism.


Regional ventricular elasticity is altered in ischemic heart disease and LV hypertrophy, potentially contributing to and sustaining a ventricular remodeling process. Conventional methods to measure regional ventricular elasticity in preclinical animal models require high fidelity ventricular pressure measurement, sonomicrometry transducers and dynamic modulation of preload. These procedures are highly invasive and provide only limited regional information; patient studies are prohibitive. We dynamically measure global LV elastance using undersampled radial real-time MRI and seek to find regional LV elastance in these studies. This requires tracking LV motion derived from wall thickness measurements in a large number of real-time image frames at high accuracy. We assessed several motion tracking algorithms and characterize their fidelity with respect to true motion labeled by experts and sensitivity to residual radial streak artifact.A series of short axis MR slices were acquired in 9 patients using a golden-angle radial bSSFP trajectory  and reconstructed via conjugate gradient-SENSE based iterative reconstruction. Two independent researchers manually segmented the datasets and the movies were restricted to three full heartbeats to limit the number of images. Image registrations were performed between the initial end-diastolic frame and all subsequent frames. The image registrations were performed using b-spline and symmetric diffeomorphic (SyN) image registration across a range of spatial scales  improved results relative to the smaller step size (0.1). We believe the ideal spatial scale is a tradeoff between sufficient resolution to capture the cardiac motion and avoiding local minimums. In terms of metrics, smaller changes in the results were observed but the mean squares resulted in the best overall result.Symmetric diffeomorphic image registration of moderate spatial scale allows for accurate tracking of myocardial motion and estimation of wall thickness from a large number of real-time image frames.K99-HL108157, R01-HL103723, T32HL007954, T32-EB009384.


Repetitive elements comprise at least 55% of the human genome with more recent estimates as high as two-thirds. Most of these elements are retrotransposons, DNA sequences that can insert copies of themselves into new genomic locations by a “copy and paste” mechanism. These mobile genetic elements play important roles in shaping genomes during evolution, and have been implicated in the etiology of many human diseases. Despite their abundance and diversity, few studies investigated the regulation of endogenous retrotransposons at the genome-wide scale, primarily because of the technical difficulties of uniquely mapping high-throughput sequencing reads to repetitive DNA.RepEnrich to study genome-wide transcriptional regulation of repetitive elements. We show that many of the Long Terminal Repeat retrotransposons in humans are transcriptionally active in a cell line-specific manner. Cancer cell lines display increased RNA Polymerase II binding to retrotransposons than cell lines derived from normal tissue. Consistent with increased transcriptional activity of retrotransposons in cancer cells we found significantly higher levels of L1 retrotransposon RNA expression in prostate tumors compared to normal-matched controls.Here we develop a new computational method called Our results support increased transcription of retrotransposons in transformed cells, which may explain the somatic retrotransposition events recently reported in several types of cancers.Supplementary material is available for this article at 10.1186/1471-2164-15-583 and is accessible for authorized users. L1PA retrotransposons are the most recent family of primate specific L1 LINE retrotransposons and show a high degree of homology due to their evolutionary recent divergence from a common L1 retrotransposon.Figure S17Effect of read length on repetitive element subfamily read assignment. Analysis of simulated data by RepEnrich reveals key features and biases associated with estimating repetitive element enrichment in high-throughput sequencing data. Read length affects the ability to distinguish between highly homologous repetitive element subfamilies. A simulated ChIP-seq sample was trimmed from 100 base pairs to 50 and 30 base pairs. The identical sample sequenced with different read lengths were analyzed by RepEnrich. The reads from each sample that were categorically assigned to repetitive elements was then examined for the number of distinctive repetitive element subfamilies the reads aligned. A) Log10 proportion of reads plotted as a function of number of repetitive element subfamilies reads aligned. B) Reads analyzed by RepEnrich were binned and determined as falling into a unique repetitive element subfamily, 2- 10 repetitive element subfamilies, or >10 subfamilies.Table S1Description of publically available datasets used in this study. Set 1–7 refers to how the datasets were grouped into separate GLM models that were used in our analysis


Delirium is a frequently occurring complication of critical care, occurring in approximately 45% of unplanned UK ICU admissions . The preThis study prospectively investigated 41 patients. Medical and surgical general ICU patients were included after 24 hours of sedation and mechanical ventilation. The researchers calculated PRE-DELIRIC scores for each patient. PRE-DELIRIC involves recording 10 variables, submitted into an online algorithm that estimates the percentage risk of delirium. We diagnosed delirium with the CAM-ICU which was performed 12 hourly [The PRE-DELIRIC scores predicted a mean rate of delirium of 39%. PRE-DELIRIC risk scores ranged from 4 to 93% Figure . Six (15Our observation that <30% of patients experienced delirium is less than the reported prevalence in similar settings and our own audits. This study demonstrates that there is some agreement between recorded rates of delirium and predicted rates using PREDELIRIC. We suggest that PRE-DELIRIC can be used in quality/audit work on UK ICUs in order to assess attempts to improve the management of delirium. Further work is required to assess the utility of PRE-DELIRIC as a risk assessment tool in individual patients.


The neurological hallmark of TTR-FAP is a length-dependent axonal neuropathy that initially involves the unmyelinated and small myelinated nerve fibers that mediate pain and temperature sensation, causing sensory disturbances that typically start in lower limbs. Subsequent degeneration of larger myelinated fibers results in large fiber sensory deficit and muscle weakness.The disease can be difficult to recognize due to extreme phenotypic heterogeneity and nonspecific clinical symptoms even within the same mutation.In TTR-FAP related to Val30Met mutation, different neuropathy phenotypes have been reported mainly in patients from non endemic areas as well in late onset cases.We described 3 TTR-FAP Val30Met early onset cases from endemic areas with a different neuropathy phenotype that can be easily misdiagnosed as a different entity. One case presented as a bilateral carpal tunnel syndrome without neuropathy; another with a demyelinating neuropathy with predominant upper limb involvement and other with a demyelinating neuropathy with conduction blocks mimicking a CIDP.TTR-FAP is frequently misdiagnosed, e.g. as idiopathic polyneuropathy or chronic inflammatory demyelinating polyneuropathy, and may be greatly under diagnosed. However, early accurate diagnosis of TTR-FAP is crucial for effective disease control.


The reverse CART technique provides the potential to modify the retrograde procedure by improving the controlled movement of the retrograde wire and improve the success rates of percutaneous coronary intervention (PCI) of chronic total occlusion (CTO). Development of interventricular hematoma is a rare complication of CTO PCI. A 63-year-old man with effort angina with a right coronary artery CTO lesion underwent PCI by retrograde approach from the LAD to a septal branch. A contrast “stain” was demonstrated surrounding the septal collateral channel used for the retrograde approach at the end of the procedure without symptom. Echocardiography indicated an increased interventricular septum thickness with low echo signals region and decreased contractility. Cardiac magnetic resonance (CMR) imaging using gadolinium showed a diffusely thickened septum with a low signal fusiform neocavitation delimited by an enhanced-signal ring suggesting intraventricular septal dissecting hematoma. After conservative treatment, follow-up echocardiogram and CMR showed the resolution of the hematoma without clinical events. This case highlights the potentially lethal complication of septal perforator dissection and hematoma that may cause severe myocardial injury caused by retrograde approach for CTO PCI. The reverse CART technique has given the potential to modify the retrograde procedure by improving the controlled movement of the retrograde wire without advancing the balloon retrogradely, which potentially reduces the septal channel injury and improves the success rates . In thisA 63-year-old man with effort angina was found to have a chronic total occlusion (CTO) of the mid right coronary artery (RCA) by coronary CT examination. Diagnostic coronary angiography confirmed the RCA CTO to be 20 mm long . RetrogrDuring the procedure, the patient remained hemodynamically stable and asymptomatic. A contrast “stain” was demonstrated surrounding the septal collateral channel used for the retrograde approach at the end of the procedure .Postprocedure echocardiography demonstrated an increased interventricular septum thickness with low echo signals core and abnormal motion . PostproDevelopment of interventricular hematoma is a rare complication of percutaneous coronary intervention for chronic total occlusion (CTO) . Previou


PET semi-quantitative methods such as relative uptake value can be robust but offer no biological information and do not account for intra-subject variability in tracer administration or clearance. Simultaneous multimodal measurements that combine PET and MRI not only permit crucial multiparametric measurements, it provides means of applying tracer kinetic modelling without the need for serial arterial blood sampling. In this study we adapted an image-derived input function (IDIF) method to improve characterization of glucose metabolism in an ongoing dementia study. Here we present preliminary results in a small group of frontotemporal dementia patients and controls. IDIF was obtained directly from dynamic PET data guided by regions of interest drawn on carotid vessels on high resolution T1-weighted MR Images. IDIF was corrected for contamination of non-arterial voxels. A validation of the method was performed in a porcine model in a PET-CT scanner comparing IDIF to direct arterial blood samples. Metabolic rate of glucose (CMRglc) was measured voxel-by-voxel in gray matter producing maps that were compared between groups. Net influx rate (Ki) and global mean CMRglc are reported. A good correlation (r = 0.9 p<0.0001) was found between corrected IDIF and input function measured from direct arterial blood sampling in the validation study. In 3 FTD and 3 controls, a trend towards hypometabolism was found in frontal, temporal and parietal lobes similar to significant differences previously reported by other groups. The global mean CMRglc and Ki observed in control subjects are in line with previous reports. In general, kinetic modelling of PET-FDG using an MR-IDIF can improve characterization of glucose metabolism in dementia. This method is feasible in multimodal studies that aim to combine PET molecular imaging with MRI as dynamic PET can be acquired along with multiple MRI measurements.


Ipilimumab blocks the immunologic checkpoint cytotoxic T lymphocyte antigen-4 (CTLA-4) and improves overall survival in patients with metastatic melanoma. Since only a subset of patients achieves benefit from ipilimumab, biomarkers that can predict patient outcome are needed. CTLA-4 blockade has been shown to affect the peripheral T cell receptor (TCR) pool, but whether diversity of the peripheral TCR repertoire is relevant as a biomarker for ipilimumab remains unknown.In this pilot study, we analyzed the baseline (pretreatment) peripheral blood TCR repertoire in 12 patients with metastatic melanoma treated with ipilimumab at 3mg/kg . TCR diversity was evaluated using a multi-N-plex polymerase chain reaction (PCR) assay which measures TCR combinatorial diversity between V and J genes from genomic DNA. The diversity of the TCR pool was studied through richness (observed V-J rearrangements) and clonality (evenness of the repertoire based on frequencies of specific V-J rearrangements). The Wilcoxon rank sum test was used to compare results between patients with clinical benefit and those without. A receiver operating characteristic (ROC) curve was used to determine an appropriate threshold for dichotomized analysis . Association with benefit in the low vs. high groups was assessed through a Fisher's exact test. Overall survival was studied through log-rank analysis.There was a significant difference in diversity richness (p = 0.033) and in clonality (p = 0.028) between patients with and without clinical benefit. Dichotomized analysis showed that none of the patients with low diversity richness  nor low clonality  achieved clinical benefit. 4 out of the 5 patients with a high clonality had clinical benefit. There was no significant difference in overall survival between patients with low vs. high diversity richness (p = 0.218) and clonality (p = 0.26).In a small group of melanoma patients treated with ipilimumab, TCR diversity in the peripheral blood at baseline was associated with patient outcomes. Further investigation is ongoing in larger cohorts of patients treated with novel immunologic checkpoint antibodies to explore these preliminary findings and determine whether TCR diversity can be used as a predictive biomarker in cancer immunotherapy.Written informed consent was obtained from the patient for publication of this abstract and any accompanying images. A copy of the written consent is available for review by the Editor of this journal.


Abnormal structural and functional plasticity in cortical and subcortical brain regions may be an important aspect of migraine pathophysiology. Resting state magnetic resonance imaging allows studying functionally interconnected brain networks. Whether there is a relation between the plasticity of resting state networks and integrity of thalamic microstructure during the migraine cycle is not known. To verify functional connectivity between brain networks at rest and its relationship with thalamic microstructure in migraine without aura (MO) patients during and between attacks.Twenty-four patients with untreated MO underwent 3T MRI scans during (n=10) or between attacks (n=14) and were compared to a group of 15 healthy volunteers. We used MRI to collect resting state data among four selected resting state networks, identified using group independent component (IC) analysis. Fractional anisotropy (FA) values of bilateral thalami were retrieved from a previous diffusion tensor imaging study on the same group of subjects and correlated with resting state ICs Z-scores.We found a significant reduced functional connectivity between the default mode network and the visuo-spatial system between attacks, and between the executive control network and the dorso-ventral attention system during attacks. When HV and migraine groups were combined, ictal and interictal selected ICs Z-scores correlated negatively with bilateral thalami FA values.The present results are the first evidence supporting the hypothesis that abnormal dynamics of the connectivity between thalamus and functional cerebral networks at rest could contribute to the recurrence of migraine attacks.Written informed consent to publication was obtained from the patient(s).


Infusions with cytokine-activated NK cells obtained from blood of MHC mismatched donors have shown some promising results especially in patients with myeloid leukaemia. There is also indirect evidence that infusions of NK cells as part of a stem cell transplant result in lower relapse rates post-transplant when donor and recipient are mismatched for KIR receptors on NK cells.Collecting NK cells form donors requires them to undergo leukapheresis with subsequent removal of CD3+ lymphocytes (to prevent GvHD) and expansion and activation of the NK cell enriched fraction with IL-2. In contrast, the continuously growing NK cell line NK-92 can be easily expanded to clinical scale in bioreactors. The broad cytotoxicity of NK-92 is due to the lack of most of the KIR receptors while expressing a range of activating receptors.We report here results from concluded and ongoing clinical phase I studies with NK-92 cells in patients with advanced cancer, that confirm its safety profile. Anti-tumour responses were seen in patients with some advanced haematological malignancies and solid tumours. NK-92 also provides a platform for further genetical engineering. A variant has been generated that expresses a high affinity FcgIIIRA receptor that can augment the treatment efficacy of mAbs that utilise ADCC to kill target cells. Various investigators are using the NK-92 cells as a platform to introduce specific tumour antigen receptors (i.e. CAR) to make them targeted to specific tumors such as melanoma, myeloma, leukaemia or brain cancer. Video-lapse studies show that those CAR engineered NK-92 cells specifically kill tumour antigen expressing cancer cells and are able to do 'serial killing'. The cell expansion process for NK-92 has been streamlined and made more economical using various bioreactor designs.The human clinical grade NK-92 cell line has many advantages over peripheral blood NK cells as a cell therapy product for cancer patients. Results from phase I clinical trials confirm its safety profile. This unique cell line is now tested in phase II studies for efficacy in various cancers and is also further developed through genetic engineering to target specific tumours through CAR. NK-92 are an off the shelf tumour targeted local and systemic cell treatment.


Ventilator-associated pneumonia (VAP) is a common and serious problem in ICUs. Several studies have been conducted to determine the effectiveness of Gram stain of tracheal aspirates for diagnosing VAP. However, the effectiveness for predicting causative microorganisms and guiding appropriate initial antibiotic therapy has not been evaluated. The purpose of this study is to determine whether Gram stain of tracheal aspirates can guide appropriate initial antibiotic therapy for VAP.We retrospectively assessed two hypothetical empirical antibiotic treatment algorithms for VAP on an 18-bed ICU. Data on consecutive episodes of microbiologically confirmed VAP were collected over a period of 22 months and divided into a derivation (1 February 2013 to 30 November 2013) and validation (1 December 2013 until 15 November 2014) cohort. We constructed two algorithms in the derivation cohort. One is a local ecology-based algorithm (LEBA), according to clinical risk factors for MDR and susceptibility results in our hospital. The other is a Gram stain-based algorithm (GSBA). The selection of antibiotics on GSBA was directed against pathogens predicted from the results of bedside Gram staining of tracheal aspirates collected just before antibiotic therapy. Subsequently, LEBA and GSBA were retrospectively reviewed and compared with actually prescribed antibiotics in the validation cohort.P < 0.001). GSBA recommended carbapenems in significantly less episodes than LEBA (P < 0.001) and the same episodes as actually prescribed initial therapy (P = 1). However, there was significant increase of antibiotic coverage rates in GSBA compared with the actually prescribed initial therapy .The first 50 VAP episodes made up the derivation cohort and the subsequent 50 VAP episodes the validation cohort. Antibiotic coverage rates by applying LEBA and GSBA were identical (96% vs. 96%). GSBA proposed more narrow spectrum therapy as compared with LEBA (Antibiotic coverage rates on GSBA were comparable with LEBA. The use of GSBA would result in a significant reduction of the administration of broad-spectrum antibiotics. Bedside Gram staining may be useful to guide appropriate initial antibiotic therapy for VAP.


Initial response to growth hormone (GH) varies considerably. In severe GH deficiency initial response varies across the group by almost 100%. In common non-GH deficient disorders such as idiopathic short stature (ISS) and children born small for gestational age (SGA) initial response varies by more than 100%.The explanation for the wide response to GH treatment can be clustered into three broad groups; (i) non-compliance with GH therapy, (ii) clinical characteristics associated with GH responsiveness and (iii) mutations and common gene variants in the GH-IGF-I axis.(i) Non-compliance with GH therapy: The commonest explanation for variable GH response is missed GH doses. In a national prospective study of GH compliance in New Zealand children; 34% of children missed more than one GH dose per week. Maori and Pacific children missed an additional GH dose each week than Caucasian children. Predictably, poor compliance (≥4 doses/week missed) was associated with a far poorer growth response than good compliance (height velocity SDS +0.4 vs +2.8). Even moderate compliance (2-3 doses/week missed) was associated with a poorer growth response (height velocity SDS +1.4 versus +2.8). Interestingly, paediatricians were unaware of GH non-compliance. GH response can only be maximised when more effective GH delivery devices are developed that record the date and time GH is administered to enable paediatricians to more effectively address compliance.(ii) Clinical characteristics associated with GH responsiveness: There is a differential response to GH therapy according to the underlying disorder. GH deficient children are more responsive than those with Turner Syndrome or idiopathic short stature who are in turn more responsive than SGA children (as shown by height velocity and serum IGF-I for a given GH dose). Furthermore, across the normal birth weight range we have recently shown that progressive reduction in birth weight SDS is associated with a progressively lower IGF-I response during IGF-I generation testing. Across growth disorders; more severe GH deficiency, taller parents, shorter stature, younger age at the start of treatment and heavier children are all parameters found to be associated with greater initial and sustained response to GH treatment. Michael Ranke has pioneered individualised GH dosing through GH prediction modelling across growth disorders. GH response can be predicted by inserting common clinical characteristics into a prediction equation that includes adjusting the dose. Prediction modelling is of greatest benefit to children with adverse clinical characteristics in whom greater initial GH doses will lead to greater growth response.(iii) Mutations and common gene variants in the GH-IGF-I axis: Downstream gene mutations in this axis  are relatively rare causes of short stature, poor growth and GH response. Conversely common gene variants are being increasingly sought that influence stature, growth and GH responsiveness. Initial reports suggested a common GH receptor exon 3 deletion was associated with a better response to GH than the full length form in SGA and ISS children. Further studies have found little if any effect of the GH receptor deletion on initial or sustained growth response in SGA, Turner Syndrome or GH deficiency. Future studies are likely to focus on panels of common gene variants across the GH axis rather than examining individual genes to better characterise GH responsiveness.


Plant responses to drought stress depend on highly regulated signal transduction pathways with multiple interactions. This complex crosstalk can lead to a physiological outcome of drought avoidance or tolerance/resistance. ABA is the principal mediator of these responses due to the regulation of stomatal closure that determines plant growth and survival, but also other strategies of droughtresistance such as osmotic adjustment. However, other hormones such as JA seem responsible for regulating a subset of plant responses to drought by regulating ABA biosynthesis and accumulation and ABA-dependent signalling, but also by ABA independent pathways. Here, we review recent reports of ABA–JA hormonal and molecular interactions within a physiological framework of drought tolerance. Understanding the physiological significance of this complex regulation offers opportunities to find strategies of drought tolerance that avoid unwanted side effects that limit growth and yield, and may allow biotechnological crop improvement. ABA is the main driver of stomatal closure which is probably the most effective mechanism to minimise dehydration. In terms of water stress sensing, stomata and roots are the most important organs that integrate environmental conditions with plant water status, with an important distinction depending on the origin (root or leaf) of the stress sensation Kramer . Both soAt the molecular level, there are thousands of ABA dependent genes controlling growth, senescence, secondary metabolism, protein biosynthesis and osmotic adjustment to coordinate plant behaviour under stress conditions  can diminish ABA accumulation  stress conditions, and is associated with resistance to biotic stress and wounding in stress signalling. Of the JA precursors, 12-oxophytodienic acid (OPDA) is one of the most important due to its possible independent roles in signalling downstream of JA. The Isoleucine conjugate form of JA, jasmonic acid Isoleucine (JA-Ile) has been characterized as the key signalling compound in JA-dependent responses due to its interaction with the receptor Coronatine-Insensitive 1 (COI1). Several recent publications have demonstrated JA’s involvement in water stress responses , NAC and MYC/MYB TFs Fig. . UsuallyOryza sativa, the MYC-family TF OsbHLH148 interacted with OsJAZs in response to drought  and jin-1  mutants showed no biomass reduction while this treatment approximately halved biomass of WT and jar1 plants  than WT plants (9 %) when water was withheld, and detached air-dried leaves had a higher photosystem II efficiency (Fv/Fm) than non-transformed plants. This suggests greater stomatal closure to maintain leaf water status and protect the photosystems. It is uncertain whether these specific mechanisms will be relevant to maintaining yields in dryland rice production. In contrast, another publication dealing with MYC2-related signalling reported that after ceasing watering, then maintaining a moderate drought by daily irrigation (leaf relative water content (RWC) averaged 65 %), both  Tardieu .versus antagonism of ABA-JA in water stress and interaction with ET) requiring caution in assessing phenotypes.The Ethylene (ET) transcription factor family are interesting targets for both JA-ABA interactions and ET. ERF5 and ERF6 are two redundant TF that are involved in leaf growth inhibition under mild drought stress  enhanced drought and cold tolerance in Arabidopsis by inducing ABA hypersensitivity  under drought (rain was excluded from 10 days before heading to 20 days after heading) and normal conditions respectively  greatly decreased grain filling rate under both normal and drought conditions. Since improved root development is a common strategy for drought avoidance; using root specific promoters . This causes ion efflux thereby lowering cellular turgor due to water efflux out of the guard cells and hence causing stomatal closure. This signal transduction pathway, along with involvement of JA and other hormones was recently reviewed  and reactive oxygen species (ROS) Fig. , due to nts Fig. . To explnts Fig. . Brieflynts Fig. . This woaba) and plants treated with the ABA biosynthesis inhibitor fluridone  induction , suggesting that MeJA treatment is JA-Ile dependent and hence COI-1 dependent. These experiments expose the variability of the stomatal responses to exogenous hormonal treatment and the necessity of assessing apoplastic hormone concentration adjacent to the guard cells to better compare epidermal bioassays with in planta experiments. This is important to distinguish hormonal crosstalk (JA influencing ABA biosynthesis in this particular example) from molecular interactions in a signalling pathway leading to a physiological outcome .However, there are also some discrepancies in the direct effect of exogenous treatment of MeJA on stomatal closure Table . In Arabpr) can also influence plant water status to mediate transpiration. While it is important not to underestimate the importance of root morphology and anatomy in mediating Lpr  had a lower Lpr  in nutrient solution can also increase Lversus JA and JA-Ile). We also need to assess the direct influence of jasmonates in responses to drought as distinct from the indirect influence through ABA biosynthesis and signalling. It is important to unveil these interactions because they offer an opportunity to exploit the natural complexity of drought signalling. This may allow us to enhance crop performance by inducing drought tolerance and resistance mechanisms that allow better yields in sub optimal conditions, thereby avoiding conservative strategies that sensitively close the stomata to maintain tissue water status, but limit photosynthesis thereby incurring yield penalties.While jasmonates seem involved in drought stress responses, a more comprehensive assessment of environmental and physiological responses  are necessary to understand the molecular mechanisms that can potentially improve crop yields in water-limited environments. Assessing ABA and JA interactions in drought stress responses needs to consider the three levels of interactions summarised in this short review . Accurate hormonal quantification of ABA and JA, JA-Ile and ideally OPDA in relevant tissues is necessary (Dave et al.


Sound-induced flash illusions(SIFI) permit to evaluate cross-modal audio-visual perception. When one flash is accompanied by two beeps, it is perceived as two flashes('fission'illusion); a 'fusion' illusion occurs when a single beep causes the fusion of a double flash stimulus. SIFI strictly depends on cortical excitability: healthy controls perceive less illusions by increasing visual cortex excitability through anodal tDCS .to evaluate if, due to cortical hyperexcitability, differences in SIFI occur in migraine and further changes can be found across migraine cycle, migraine chronification an drug overuse.we enrolled 64 patients with episodic migraine, 32 with-(MWA) and 32 without-aura(MWO) , 44 patients with chronic migraine with medication overuse headache , and 20 healthy controls . All underwent a paradigm for SIFI induction where had to report the number of flashes seen. 13 of MWO and 12 out of MWA were examined in both ictal and interictal phaseall migraine groups showed significantly less SIFI than controls(p<.0001); illusions are more reduced in in chronic migraine and particularly in those overusing triptans(p<.05).results point to a condition of visual cortical hyperresponsivity in patients with chronic migraine in analogy to what observed in episodic patients expecially during ictal phase. This is in agreement with the view of chronic migraine as a 'never ending attack'. The greater effect showed in triptan overuser can follow to down-regulation of 5HT1 receptors.No conflict of interest.


To communicate imaging findings and interpretation precisely and comprehensively within a multidisciplinary treatment team written radiology reports are essential. Structured reporting systems increasingly take advantage of conventional free-form reports. Therefore we developed a visual imaging report chart and evaluated its acceptance by radiologists and oncologists in comprehensive cancer centers.Six imaging report charts of representative CT follow-up examination of patients with metastatic melanoma were recorded. Each chart includes a short and focused written text as well as a visual reporting diagram, which is composed of two sections: (1) patient characteristics; and (2) schematic documentation and graphical and pictorial visualization of the radiologic findings including documentation of imaging findings, treatment response according to qualitative or standardised criteria, clinical recommendation and the course of tumour burden. In total 36 reviewers  compared the report charts with the corresponding conventional free-text report by a questionnaire grading their degree of satisfaction by a 10 point Likert-scale. Statistical significance was evaluated by one-way analysis of variance (ANOVA) and Student’s t-test.Referring oncologists rated improvement of therapy decision making for oncologic patients significantly higher than radiologists (p<0.01). A significant difference (p<0.005) was observed between radiologists and clinicians regarding their opinion on time-saving in working routine due to our reporting chart (5.1 vs. 8.7 respectively).Structured reporting and documentation of CT examinations in metastatic disease is well appreciated by referring oncologists. The proposed standardised imaging report chart has the potential to improve therapy decision making in the multidisciplinary context.


Current commercial wall linings are produced using labour intensive fabric weaving methods. This study explored the effectiveness of inexpensive monofilament mesh linings produced by extruding insecticide impregnated polyethylene and poly(ethylene-co-vinyl acetate) (EVA) directly into a net format.Dichlorodiphenyltrichloroethane (DDT), pyrethroids, organophosphates and carbamates are insecticides recommended by the World Health Organisation (WHO) for malaria vector control . Long LiPolyethylene and EVA were extruded with pyrethroids  and organophosphates (ca. 0.5 wt.% insecticide) respectively through a dye to form monofilament meshes. Scanning Electron Microscopy (SEM) and Confocal Fluorescence Microscopy (CFM) was used to characterize the presence of insecticide on the surface of the monofilaments. Efficacy testing was done using standard WHO bioassay tube tests.® to be effective, the insecticides have to be available on the monofilament surface since they are contact poisons. SEM and CFM micrographs show that pyrethroids are present and evenly distributed on the surface of the monofilament mesh. The WHO effectiveness criterion is a mortality exceeding 80% after 24 hours following a 30 minute exposure of An. arabiensis mosquitoes to test samples. The pyrethroid impregnated mesh samples achieved 100% mortality in this test even after 24 months of storage. Samples impregnated with organophosphates failed to achieve the WHO criterion. Poor performance of the organophosphate insecticides in the EVA matrix was probably due to the high solubility of these insecticides in this polymer.For NetlonMonofilament polyethylene mesh impregnated with pyrethroid insecticides was successfully produced by a simple direct extrusion technique. It is an inexpensive method for the production of slow-release polymer-based wall linings. Laboratory efficacy tests suggest that this ITWL technology may be a potential substitute for IRS and it may also complement LLIN.


Durable complete response rates of 20% have been observed in clinical trials of patients with metastatic melanoma employing adoptive cell transfer (ACT) of patient derived tumor infiltrating lymphocytes (TIL). Here we provide a detailed analysis of the response of TIL administered to a patient with metastatic melanoma who exhibited a complete response ongoing greater than 3 years. Using whole exome and RNA sequencing and bioinformatic analysis of the patient's matched tumor and normal gDNA we identified over 4,000 non-synonymous somatic mutation variants. We screened 745 somatically mutated genes using tandem minigene constructs expressing transcripts expressed in autologous tumor cells whose expression levels were greater than 0.1% of the levels of β- actin. These tandem minigenes were then transfected into autologous B cells and then analyzed for their ability to stimulate the administered T cells. Our results indicated that the autologous TIL distinctly recognized 10 somatically mutated gene products, each of which was recognized in the context of three different HLA class I restriction elements expressed by the patient's tumor. Detailed T cell clonal analysis revealed that 9 of the top 20 most prevalent clones present in the infused TIL, comprised over ¼ of total infused cells and recognized mutated antigens. These results further supported our efforts to identify and enrich mutation-reactive T cells for the treatment of patients with metastatic cancer.


Telepathology practice across international borders has become increasingly popular. Two years after the University of Pittsburgh Medical Center (UPMC) launched a telepathology consultation service with KingMed Diagnostics Laboratory in Guangzhou, China, latency issues occurred when viewing whole slide images (WSI). Our aim was to explore various image file management solutions to improve the viewing experience of digital consult cases.WSI files generated by KingMed when scanning glass slides for consultation initially resided on a Hamamatsu server, using the NDP.serve database management system. These remote files were securely accessed using a custom portal via the Internet. To try solving ongoing latency issues, WSI files were instead transferred from China to the UPMC data center using an open source product . This was a command line utility placed into a batch process. A faster high speed file transfer software solution (Aspera) was subsequently employed. This commercial software allowed immediate file transfers to occur without a user initiating the transfer.Viewing digital consult cases from Pittsburgh in the USA with WSI files residing on a server in China negatively impacted viewing of images. Image display deteriorated to about 2 minutes/case. Transferring files with the open source product provided transfer speeds of 2-3 MB/second but suffered from intermittent dropped connections. Employing the commercial software permitted more reliable transmission of digital files with 75–100 MB/second transfer speeds.Successful global telepathology requires dedicated image management. Transfer of files to local servers delayed the process by up to 24 hours but greatly improved the overall turn-around time of digital consultations. This was partly negated by employing high speed file transfer software. Transfer of digital files helped overcome network latency issues experienced with China and enhanced the viewing experience for end-user digital consultants.


This study investigated the safety profile and potential "therapeutic" effect of intravenous ultrasmall superparamagnetic iron-oxide (USPIO)-based iron administration regarding infarct healing in patients with ST-elevation myocardial infarction (STEMI).Seventeen study patients  and 22 matched controls  both with primary reperfused STEMI underwent multi-parametric CMR studies in the first week and three months after acute MI. Only IRON patients received a single intravenous bolus of 510 mg elemental iron as ferumoxytol (FerahemeTM) within four days following acute MI.Three months after study inclusion, all patients were alive and there were no adverse cardiac events. Significant improvement in left ventricular (LV) ejection fraction  as well as shrinkage of infarct size were seen in both groups at follow-up. There was a more pronounced decrease in infarct size in the IRON group  in addition to a significant decrease in both endocardial extent and prevalence of transmural infarctions in IRON but not in CONTROL patients. A significant decrease in LV end systolic volume was only seen in the IRON group .Intravenous iron administration in acute STEMI patients is associated with both improved infarct healing and beneficial global left ventricular remodelling (compared to matched controls). These findings together with the good safety profile make USPIO-based iron administration a promising future candidate as a "diagnostic" and "therapeutic" adjunctive solution in acute MI management.This work was financially supported by a grant from the German Federal Ministry of Education and Research .


Vascular cerebral dysfunction restricted to the posterior circulation in migraineurs has been reported. In this preliminary study we compared mean flow diastolic velocity in the posterior cerebral artery during visual stimulation in interictal state headache-free subjects and migraineurs with aura.Trancranial color-coded duplexsonography was used to assess mean flow diastolic and systolic velocity changes in the posterior cerebral artery, in particular P2, during visual stimulation.An increase in vascular reactivity of mean flow diastolic velocity was observed during visual stimulation in headache-free subjects  and in the patients affected by migraine with aura , without any differences between right and left sides. Reactivity of mean flow diastolic velocity to visual stimulation was found significantly higher in Group 2  vs Group 2  (p < 0.001). In contrast, no differences in flow systolic velocity between the two groups were observed.These results indicate that occipital cortex is involved in migraineur patients with aura. Moreover, these patients exhibit a larger cerebrovascular response during visual stimulation compared to headache-free subjects.Written informed consent to publish was obtained from the patient(s).


Managing spinal deformities in young children is challenging, particularly early onset scoliosis (EOS). Surgical intervention is often required if EOS has been unresponsive to conservative treatment particularly with rapidly progressive curves. An emerging treatment option for EOS is fusionless scoliosis surgery. Similar to bracing, this surgical option potentially harnesses growth, motion and function of the spine along with correcting spinal deformity. Dual growing rods are one such fusionless treatment, which aims to modulate growth of the vertebrae. The aim of this study was to ascertain the extent to which semi-constrained growing rods (Medtronic) with a telescopic sleeve component, reduce rotational constraint on the spine compared with standard rigid rods and hence potentially provide a more physiological mechanical environment for the growing spine.Six 40-60kg English Large White porcine spines served as a model for the paediatric human spine. Each spine was dissected into a 7 level thoracolumbar multi-segment unit (MSU), removing all non-ligamentous soft tissues and leaving 3cm of ribs either side. Pure nondestructive axial rotation moments of +/-4Nm at a constant rotation rate of 8deg/s were applied to the mounted MSU spines using a biaxial Instron testing machine. Displacement of each vertebral level was captured using a 3D motion tracking system (Optotrak). Each spine was tested in an un-instrumented state first and then with appropriately sized semi-constrained growing rods and rigid rods in alternating sequence. The rods were secured by multi-axial pedicle screws (Medtronic) at levels 2 and 6 of the construct. The range of motion (ROM), neutral zone (NZ) size and stiffness (Nm/deg) were calculated from the Instron load-displacement data and intervertebral ROM was calculated through a MATLAB algorithm from Optotrak data.Irrespective of the order of testing, rigid rods significantly reduced the total ROM compared with semi-constrained rods (p<0.05) which resulted in a significantly stiffer spine for both left and right axial rotation (p<0.05). Analysing the intervertebral motion within the instrumented levels 2-6, rigid rods showed reduced ROM compared with semi-constrained growing rods and compared with un-instrumented motion segments.Semi-constrained growing rods maintain similar stiffness in axial rotation to un-instrumented spines, while dual rigid rods significantly reduce axial rotation. Clinically the effect of semi-constrained growing rods as observed in this study is that they would be expected to allow growth via the telescopic rod components while maintaining the axial flexibility of the spine and improved capacity for final correction.


A 53-year-old man, previously healthy, presented with a slow-progressive enlarging mass in the midfrontal area which had developed 14 years previously without headache. On physical examination, this nodular reddish mass (11 x 7 x 5 cm) was fluctuant to pressure and tethered to the underlying frontal bone (A). There was no neurological deficit and no superficial lymph nodes. Cranial computed tomography scan showed an irregular median calvarial defect with poorly bony defined margins and left frontal sinus invasion (B-C). Magnetic resonance imaging revealed intracranial extension of the tumour which invaded the dura and the superior longitudinal sinus (D-E). Local biopsy was performed and histological study consisted with a dermatofibrosarcoma. In this specific case, radiotherapy was planned before surgical excision and craniofacial reconstruction. Dermatofibrosarcoma is a slow-growing mesenchymatous tumour of the skin with high local malignancy and great opportunity of recurrence. It usually occurs in the trunk or extremities. In the majority of cases the tumour remains asymptomatic for a long time. Head  and maxillofacial involvement is rare particularly with skull vault and intracranial extension. To improve local control after surgery, wide excision is recommended but difficult in significant anatomic region as in our case.


It is unclear if children after heart transplantation (HTX) are at risk for developing increased myocardial fibrosis. Diffuse myocardial fibrosis can be estimated by myocardial longitudinal relaxation (T1) times.Twenty cardiovascular magnetic resonance (CMR) studies in 17 patients after HTX  were analysed retrospectively and compared to CMR studies in nine healthy controls . Patients with clinically significant rejection were excluded. T1 measurements were performed at a single mid-ventricular short axis slice orientation before and > 10 minutes after the application of 0.2 mmol/kg gadopentetate dimeglumine (Gd) in the interventricular septum (IVS), LV lateral wall and the complete LV myocardium (Image). The tissue-blood partition coefficient was calculated as a function of the ratio of T1 change of myocardium as compared to blood.Pre-contrast T1 times before the application of Gd were significantly higher in HTX patients compared to controls , whereas the reduced post-contrast T1 times in the HTX patients showed a trend towards being shorter than in controls but failed to reach statistical significance. Tissue-blood partition coefficients were elevated in patients after HTX in the LV lateral wall  and the complete LV myocardium . The difference in the IVS failed to reach statistical significance .Diffuse fibrosis is present in children after HTX as evidenced by pre- and post-contrast myocardial T1 mapping. The technique may be suitable for the detection of early signs of adverse remodeling after HTX.This study was partly funded by the Labatt Family Heart Centre, Witchell Fellowship, and by Siemens.


Negr1 gene locus retrieved from the UCSC genome browser. This approach could be adopted to identify putative regulatory non-coding RNAs in other tissues and diseases.Long non-coding RNAs and microRNAs control gene expression to determine central nervous system development and function. Neuronal growth regulator 1 (NEGR1) is a cell adhesion molecule that plays an important role in neurite outgrowth during neuronal development and its precise expression is crucial for correct brain development. The data described here is related to the research article titled “A long non-coding RNA, BC048612 and a microRNA, miR-203 coordinate the gene expression of Neuronal growth regulator 1 (NEGR1) adhesion protein”  Specifications TableValue of the data•in silico analysis of the genetic signatures and expression pattern of long non-coding RNAs associated with the Negr1 gene.This data provides a comprehensive •Negr1 gene expression.This data is useful in understanding the regulatory relationship between non-coding RNA gene expression and •The methodology provided can be used to postulate regulatory relationships between long non-coding RNAs and proximal genes.•This approach could be adopted to identify putative regulatory non-coding RNAs in other tissues and diseases which could be evaluated with further functional studies.in silico analysis carried out on the Negr1 gene locus for genetic signatures and expression patterns of long non-coding RNAs in proximity to the Negr1 gene The data in this article describes the initial Negr1 gene promoter region was analyzed using the UCSC Genome Browser Negr1 gene promoter  The promoter . PresencNegr1 gene transcription start sites as well as putative regulatory transcription factors was carried out (Further interrogation of the BC048612 lncRNA and ried out . RegRNA ried out .To determine if there is any interaction between the BC048612 lncRNA and miR-203, the BC048612 sequence was analyzed for putative binding sites for miR-203. RegRNA analysis did not predict any miR-203 binding sites on the BC048612 lncRNA The authors declare that they have no competing interests.


Insects and their six-legged relatives (Hexapoda) comprise more than half of all described species and dominate terrestrial and freshwater ecosystems. Understanding the macroevolutionary processes generating this richness requires a historical perspective, but the fossil record of hexapods is patchy and incomplete. Dated molecular phylogenies provide an alternative perspective on divergence times and have been combined with birth-death models to infer patterns of diversification across a range of taxonomic groups. Here we generate a dated phylogeny of hexapod families, based on previously published sequence data and literature derived constraints, in order to identify the broad pattern of macroevolutionary changes responsible for the composition of the extant hexapod fauna. The most prominent increase in diversification identified is associated with the origin of complete metamorphosis, confirming this as a key innovation in promoting insect diversity. Subsequent reductions are recovered for several groups previously identified as having a higher fossil diversity during the Mesozoic. In addition, a number of recently derived taxa are found to have radiated following the development of flowering plant (angiosperm) floras during the mid-Cretaceous. These results reveal that the composition of the modern hexapod fauna is a product of a key developmental innovation, combined with multiple and varied evolutionary responses to environmental changes from the mid Cretaceous floral transition onward. Hexapoda, including insects and their six-legged relatives, are the most species-rich animal clade in terrestrial ecosystems and collectively comprise over half of all described extant species Attempts to explicitly test these ideas within a phylogenetic framework have either been restricted to particular orders The dated phylogeny used in this study contains 874 higher taxa of Hexapoda . Taxa weRhyniognatha hirsti from the early Devonian Using our dated tree we estimated the crown divergence of Hexapoda, i.e. the divergence of true insects from Entognatha  as occurring in the Ordovician , which is consistent with other recent molecular clock estimates At higher taxonomic levels, lineage through time plots  indicateDespite this apparent stability in the origination of higher taxa, the application of birth-death models In addition to these broad patterns, diversification shift models identified a further forty-three clades on the tree potentially associated with shifts in diversification rates . These sIn contrast with these relict groups, most of the shifts leading to a net increase in taxonomic richness are comparatively recent  and are Unsurprisingly, several upshifts in diversification within Holometabola also involve groups directly associated with the angiosperm radiation, with notable examples including leaf and longhorn beetles (Chrysomeloidea) Ultimate explanations of insect diversification can be classified into morphological key innovations and ecological interactions The phylogeny of 874 terminal taxa, representing familial groups within Hexapoda was inferred based on eight widely sampled molecular markers including nuclear  and mitochondrial  protein coding sequences and 16S, 18S and 28S rRNA sequences. All included taxa are chimeric, i.e. the sequences used are assembled from multiple individuals and species, such as to reduce the problem of non-overlapping sampling within the source datasets and to maximize gene coverage for each of the sampled terminals. All gene partitions were aligned using MAFFT The fully resolved maximum likelihood topology estimated above was used as the basis for a relaxed independent gamma rates clock implemented in Mr. Bayes 3.2 Estimates of extant richness for terminal taxa were compiled from recent encyclopedic sources . Models of clade diversification were implemented in R v 2.15.1 Figure S1Nodal support on the phylogeny. Nodes marked with circles are either constrained (red) or have high bootstrap support .(TIF)Click here for additional data file.Figure S2Topology showing 95% confidence intervals on node ages (transparent blue bars). Black rings denote 100 Ma intervals from the present. Nodes denoted with red circles are involved in calibration (see (TIF)Click here for additional data file.Figure S3The fifty shifts with the highest rates of recovery in samples from the MCMC chain  plotted  Shifts are denoted as (TIF)Click here for additional data file.Table S1Estimates of extant species richness for terminal groups and GenBank accession numbers for sequences used in phylogenetic reconstruction.(XLSX)Click here for additional data file.Table S2Fossil calibrations implemented in dating the tree topology. Calibrated nodes are plotted on (XLSX)Click here for additional data file.Table S3Parameter values and shifts in species richness associated with MEDUSA model shifts inferred across the mean topology. Shifts listed here are plotted on (XLSX)Click here for additional data file.Table S4The fifty most robustly recovered shifts inferred from 500 samples from the post-burnin Markov Chain Monte Carlo (MCMC). Shifts are plotted on (XLSX)Click here for additional data file.Datafile S1Tree topology in Nexus format. Contains two tree files with a) an undated cladogram including bootstrap support for nodes and b) a dated topology including confidence intervals for the node ages and denotation of inferred shift events matching (TXT)Click here for additional data file.Datafile S2Alignment and MrBayes instructions in Nexus format. Contains the implemented alignment after processing with the commands and priors used in setting the MrBayes dating run.(TXT)Click here for additional data file.Text S1Supplementary Experimental Procedures and Discussion. Contains further details of experimental procedures, discussion of topology and reliability of diversification shift estimates, and cited references for fossil calibrations and species richness estimates Click here for additional data file.


Idiopathic inflammatory myopathies are heterogeneous systemic autoimmune diseases that are characterized by progressive muscle weakness and typical inflammatory infiltration in skeletal muscle. It is known that a cardiac involvement may occur; the frequency varies between 9% and 72%, mainly depending on patient selection and diagnostic procedures applied. Most common finding is a progressive (systolic) heart failure. Diagnosis may be difficult in patients that exhibit exercise limitation due to peripheral muscle weakness.It has been shown that cardiac magnetic resonance imaging (CMR) is reliably able to provide important information concerning cardiac function and inflammatory processes in a single exam. Therefore, objective of our prospective study was to assess and characterize myocardial involvement in patients with histological defined idiopathic inflammatory myopathies by a comprehensive CMR protocol.Fifty-three consecutive patients with histologically proven idiopathic inflammatory myopathy were enrolled into the study.All patients underwent CMR examination in a 1.5-T whole body scanner using a 32-channel phased-array cardiac surface coil. Volumetric and functional assessment was performed using 3D short-axis steady-state free precession sequences covering the entire left and right ventricle. Early myocardial enhancement was assessed before and shortly after the admission of gadolinium based contrast agent by T1-weighted fast spin-echo sequences. Inversion-recovery gradient-echo sequences were acquired for the evaluation of late gadolinium enhancement (LGE) 10-15 minutes after administration of 0.2 mmol/kg gadolinium based contrast agent .CMR images were analyzed by two experienced and blinded readers in consensus.Reduced left ventricular function (ejection fraction < 60%) was present in 9 (17%) patients. These patients exhibited significantly higher blood levels of CK-MB  than patients with preserved LVEF. Early myocardial enhancement  was more often present  as well as LGE patterns consistent with myocardial inflammation . LGE was most often observed in the inferior and lateral segments.The results of this study show that CMR is able to detect cardiac involvement in idiopathic inflammatory myopathies. Early diagnosis using CMR could result in early therapy of heart failure, leading to beneficial ventricular remodeling and thus improving quality of life and survival in these patients.


Invasive glioblastoma multiforme (GBM) is the most common malignant primary tumor of the central nervous system with an estimated annual incidence of 3.15 cases per 100,000 in the USA. Current therapies have been unsuccessful in curing malignant gliomas that are highly proliferative and infiltrating, containing regions of necrosis and hypervascularization. Novel therapeutic approaches are critically needed. A glioma specific immune response offers a potential anti-tumor benefit by immune targeted elimination of tumor cells and memory capability that may prevent future recurrence, a feature of aggressive GBM. A number of GBM tumor associated antigens (TAAs) have been studied in preclinical and clinical studies. However, most of these TAAs are HLA motif predicted T cell epitopes derived from over-expressed proteins. A systematic analysis of the naturally processed and therefore most clinically relevant MHC class I associated T cell epitopes presented by glioma cells has never been reported. A number of tumor associated over expressed glycoproteins have been identified as biomarkers and have been proposed as a potential immunotherapeutic target for various cancers including GBM. Tumorigenesis involves dysregulation such as over production or high turnover of proteins that are involved in tumor initiation, progression, unlimited growth, invasion, immune evasion, therapy resistance and survival. One of the key features of dysregulation is aberrant glycosylation of glycoproteins that are involved in these cancer pathways. In this study, we attempt to identify and characterize T cell epitopes presented on the U87 human glioblastoma cells and select epitopes that are derived from glycosylated proteins to develop a multi-epitope based therapeutic vaccine product that induce specific T cell responses against lethal GBM. Towards this goal, we enriched GlcNAc expressing glycoproteins using L-PHA lectin and identified total of 66 glycopeptides derived from 59 different glycoproteins with aberrant GlcNAc glycosylation from U87 cells via glycoproteomic analysis. We also used an immunoproteomic approach to investigate MHC class I presented peptides in tumor cells. Those selected MHC class I epitiopes were derived from glycosylated proteins that had been identified previously. In addition, these peptides were able to activate CD8+ T cell responses against glioma cells. Thus the selection of T cell epitopes derived from aberrantly glycosylated proteins forms the foundation for the development of a multi-epitope based therapeutic vaccine that induces specific T cell responses against lethal GBM.


Molecular chaperone Hsp70 is well known for its ability to stimulate innate and adaptive anti-cancer immune response. In in vivo studies we proved the efficacy of the intratumoral delivery of Hsp70 in the model of intracranial glioma C6 in animals. Based on our preclinical 12 patients with diagnosis of malignant brain tumour were enrolled in clinical trial. Following tumour resection protein was infused into tumor cavity. Specific immune response was evaluated in delayed hypersensitivity test (DTH). Peripheral blood was monitored for possible changes in lymphocyte subpopulations cytokine levels and cytolityc activity of NK-cells. Additionally cytokine production was analyzed in dynamics in liquor after injection of Hsp70. All patients were monitored for adverse effects.2-helpers towards cytokines from Th1-mediated response . This data corresponded to changes in lymphocyte subpopulations. NK-cell activity was not altered.Intratumoral injections of Hsp70 were well tolerated in patients. One patient had complete clinical response and one partial response documented by radiological findings. In three patients we observed positive DTH-test. In peripheral blood we observed a shift from cytokines of those provided by ThThis pilot study demonstrated for the first time feasibility and safety of intratumoral delivery of recombinant Hsp70 in cancer patients. Based on our preclinical and clinical findings we proposed the novel hypothesis of immunomodulatory mechanism of Hsp70. Our results suggest that purified Hsp70 can induce specific effective anti-tumor immune response and warrants further investigation in randomised clinical trials.


EditorialAcquired Immune Deficiency Syndrome (AIDS) is a social-health problem. Injection drug use, sharing needles and sexual contacts are considered as the main ways of Human Immunodeficiency Virus (HIV) transmission from person to person in Central Asian countries as well as Iran . In rece


Cardiac stress MRI has grown to a well established method to detect hypokinesia or perfusion deficits due to ischemia. Commonly, myocardial load is generated by the administration of adenosine or dobutamine, which is associated by a higher complication rate compared to exercise stress testing. Our purpose was to evaluate the use of an MR conditional pedal ergometer for cardiac stress MRI.We included 15 patients with coronary heart disease until now. All patients underwent exercise stress MRI at a whole-body 3 Tesla MR system and received real-time TRUFI CINE imaging and first-pass perfusion imaging before and after stress testing as well as late enhancement (LE) PSIR sequences after stress. All images were visually rated by three experienced radiologists.As shown on LE images, two patients suffered from silent myocardial infarction and showed stress-induced hypokinesia and slight perfusion deficits nearby the infarction scar. In additional 5 patients, we detected segmental hypokinesia and endocardial perfusion deficits due to exercise. 3 patients exhibited endocardial perfusion deficits without hypokinesia. 5 patient did not show any abnormalities due to stress.Exercise stress of the myocardium is a helpful method to detect ischemic area in cardiac MRI.Nothing to declare.


A 21 year-old male patient was admitted with symptoms of exertional dyspnea andpalpitation. The physical examination revealed a grade 3/6 systolic murmur, best heardover the left 3-4th intercostal space. Transthoracic echocardiography disclosed aseparate chamber (asterisk) in the interventricular septum. The apical portion of thechamber consisted of muscle tissue and the basal portion consisted of membranous andaneurysmatic tissue (Panel A). There was a muscular "tunnel-like" structure connectingthe left ventricle and chamber at mid ventricular level. The color and continuous waveDoppler imaging revealed a bidirectional flow across the passage .The patient underwent three dimensional transthoracic echocardiographic examinations,which revealed two separate septa. Between these septa there was a third chamber(asterisk). It was connected to both left (via the tunnel at the muscular septum) andright ventricles . The membranousseptal aneurysm was separated from the left ventricle by a thin membrane, without anypassage across it (Panel D). MRI findings were consistent with echocardiographic images(Panel E). Ventriculography and coronary angiography were performed. The coronaryarteries were normal; the left ventriculography showed the muscular andmembranous/aneurysmatic portions of the malformation (Panel F). The patient was advisedto undergo a surgical procedure, but he refused and was discharged with recommendationsabout control visits.


Right ventricular (RV) dilatation is a key imaging criterion of the 2010 modified task force criteria for the diagnosis of arrhythmogenic right ventricular cardiomyopathy/dysplasia (ARVC/D), which makes accurate and reproducible volumetric anaylsis essential. We wanted to evaluate the effect of using new thresholding software in volume analysis compared to standard manual contouring on reproducibility and accuracy.Retrospective RV volume analysis was performed on 20 anonymised cases of suspected ARVC. 8mm contiguous short-axis cine steady-state free precession (SSFP) data was acquired from the base of the ventricles to the apex on a 1.5 Tesla MRI scanner. Volumetric analysis was then analysed using two different commercially available softwares: (1) Semi-automated blood pool thresholding with CVI42  and (2) Manual contouring with Argus Syngo . Independent analysis was performed by four different experienced observers, two for each software. RV papilla and trabeculations were excluded from the blood volume. Indexed RV end diastolic volume (IEDV), end systolic volume (IESV) and ejection fraction (EF) were recorded. Statistical analysis included Bland-Altman plots and intraclass correlation coefficients (ICC) to assess for accuracy and interobserver variability.There was excellent interobserver agreement in RV volumes for both blood pool thresholding  and manual contouring , with no statistically significant difference between each set of two observers (p>0.05).2=0.93, IESV r2=0.95), Bland-Altman analysis demonstrated significant mean bias . The thresholding values were consistently lower than the manual contouring.Whilst there was good correlation between indexed volumes between the two methods (IEDV rThe average contouring time with thresholding was slightly shorter at 190 seconds compared to 262 seconds for manual contouring.The significant mean bias between thresholding and manual contouring raises important questions about the reliability between different tools we use to perform volume analysis. The bias may due to enhanced detection of more subtle trabeculations in the RV wall which would normally be excluded manually by the observer. Partial voluming occurs at the interface of the endocardium and blood pool, which may cause thresholding software to underestimate volumes. In addition, it is unclear if the software used itself may contribute to this bias. This is of particular importance in ARVC where there are currently clearly defined thresholds for abnormal RV volumes, however these factors should be considered whenever instutions upgrade software, or when comparisons are made to reference values performed with specific software.This research was supported by NIHR Bristol Cardiovascular Biomedical Research Unit.


Evidence of abnormal color vision processing in migraine comes from observation of positive symptoms during visual aura, effects of strong color contrast triggering attacks and of colored-spectacles reducing migraine frequency. Although the central or peripheral basis of such color misperception remains unclear, several authors reported a selective deficit of shortwavelength cones (S-cones). Sound-iTwenty-two MoA patients  and 12 unimpaired healthy volunteers with no family history of migraine  were enrolled. Migraine patients were tested interictally. None of the patients enrolled had taken any prophylactic drug during the 3 months prior to the experiment.Every participant took part in two randomized experimental sessions. Experiment 1 consisted of the Farnsworth-Munsell 100-hue test: the disks in 4 different trays had to be arranged to form a smooth color sequence between two reference disks. Experiment 2 consisted in reporting the number of flashes seen on a black screen in isolation or in different combinations with beeps.Cone deficits were more prevalent in the migraine (72%) than in the control group (0.08%). S-cone deficit (56.2%) was still the most frequent alteration detected in our sample. Accordingly with previous results MoA patients as a whole showed no significant difference in SIFI perception as compared to controls even if a trend towards reduced illusory percept was found. However, when performing subgroup analyses, the MoA group with S-Cone defect showed significantly less fission illusions as compared to the group of healthy subjects (p < .0001).S-Cone dysfunction is highly prevalent in migraine; patients with such functional retinal impairment show higher levels of visual cortical excitability. The results of the study seem to establish a strong  relationship between retinal and visual cortical dysfunctions found in migraine. Further evidence  is needed to define the role and interplay of such abnormalities in migraine pathophysiology.Written informed consent to publication was obtained from the patient(s).


Hybrid surgical approaches are gaining popularity. Minimally invasive aortic valve replacement and percutaneous coronary intervention [PCI] is well documented. We wanted to explore the feasibility of trans-catheter aortic valve implantation [TAVI] and off pump coronary artery bypass grafts [OPCAB].We proposed that in TAVI patients where PCI is not possible or has failed, trans-aortic TAVI with concomitant OPCAB would be a suitable alternative.A 77 year old man admitted following an out of hospital cardiac arrest was found to have an occluded left anterior descending (LAD) artery, a patent stent in a dominant circumflex and a small diseased right coronary. Past medical history included myocardial infarctions on four occasions, PCI, and severe left ventricular failure. Echocardiography demonstrated severe, calcific aortic stenosis and poor left ventricular function. The multi-disciplinary team recommended TAVI. Work up identified abdominal aortic aneurysm which precluded trans-femoral TAVI approach. A perfusion scan confirmed viable myocardium in the LAD territory. He was put forward for trans-aortic TAVI with OPCAB surgery .The procedure was performed as planned without any complications. Post-operative echocardiogram confirmed a well-placed valve. The patient was treated for a chest infection and discharged home on 8th post-operative day. He was seen at the outpatient clinic at 8 weeks and was asymptomatic and with good exercise toleranceTrans-aortic TAVI and OPCAB is a potential alternative to conventional aortic valve replacement and coronary artery bypass surgery as well as the hybrid approach of minimally invasive aortic valve replacement and PCI.Written informed consent was obtained from the patient's next of kin for publication of this abstract and any accompanying images. A copy of the written consent is available for review by the Editor of this journal.


Weaning from mechanical ventilation is an important concern in ICU clinical practice. Surface electromyography (sEMG)  is a nonTwenty-one adult and clinically stable patients undergoing assisted mechanical ventilation for more than 48 hours were investigated during pressure support (baseline) and during a 2-hour CPAP trial. sEMG of diaphragm (costmar), intercostal and sternocleidal (accessory muscles) was recorded with a dedicated device  simultaneously with airway waveforms and expressed as the ratio of the signal during baseline. Diaphragmatic electrical activity from a nasogastric tube (EAdi) of 14 of those patients was also measured.P = 0.7). On the other hand, the activation of accessory muscles increased significantly during the same period (P = 0.01) and was strongly correlated with respiratory rate  and inversely with tidal volume . In patients with EAdi we confirmed a tight correlation between costmar and EAdi . See Figure The rapid shallow breathing index was lower than 105 in all patients and only one patient failed the trial. We observed that the mean inspiratory value of costmar increased immediately after switch to CPAP but did not significantly vary during the CPAP trial (ANOVA, sEMG indicated that while diaphragm activation remains constant during the CPAP period, accessory muscles were progressively recruited and particularly in the conditions of increased respiratory rate and lower tidal volumes.


The promyelocytic leukemia zinc finger (PLZF) transcription factor is directly implicated in tumor suppression although the relevant target genes remain poorly defined. Shi et al. now implicate smooth muscle α-actin and changes in cytoskeletal architecture as key downstream targets of PLZF in opposing cellular transformation.PLZF is a transcriptional repressor belonging to the POZ-Krüppel (POK) family of transcription factors with critical roles in oncogenesis, development and stem cell maintenance -7. The aSubsequent to its original characterization in the context of leukemia development, tumor suppressive functions have been attributed to PLZF in other cell types and tissues -19. UsinOncogenic stimuli that drive cellular transformation often trigger an accompanying remodeling of the cytoskeleton that results in altered cell morphology and growth properties . Further


Monitoring guidance for clinical trials recommends the use of central and on-site monitoring practices. Comparing the efficiency of on-site monitoring visits targeted as a result of central statistical monitoring (CSM) procedures, versus those that were not, could help to improve trial conduct.The monitoring plan in a large long-term multicentre international trial required that sites received routine monitoring visits every nine months. Oversight of this trial was augmented by central statistical monitoring that identified high scoring sites as priority for further investigation. To validate this approach high scoring sites, and some low scoring sites (in the same countries) identified by the country teams as potentially problematic were visited.Over a twelve month period 21 sites  received a comprehensive monitoring visit from the senior monitor. Only 1 site identified by CSM had no findings, versus 7 of the comparator sites. Minor findings indicative of ‘sloppy practice’ were identified at 12 sites (10 CSM versus 2 comparator). At 1 site identified by CSM there were serious findings indicative of an under-performing site.On-site visits triggered by CSM are more efficient than undirected routine visits. Furthermore, information from CSM can help focus the nature of on-site visits and any interventions required to improve site quality, e.g. additional training on data collection or event reporting. Study costs will be reduced if resources are directed to improving the performance of sites of concern, leaving lower risk sites to receive less frequent on-site monitoring.


High Flow Nasal Cannulae (HNFC) is commonly used to improve oxygenation in critically ill patients. But it effects on diaphragm motion and end expiratory lung volumes (EELV) are not well described.To assess and compare effects of different flows of HFNC and conventional facemask on the diaphragm excursion. Secondary outcomes were to assess effects on patient discomfort, PaO2/FiO2 ratio, EELI and TDI variation measured by electrical impedance tomography (EIT).Cross-over single prospective study on patients undergoing a major abdominal surgery and high respiratory risk assessed by ARISCAT score > 26 admitted in a surgical intensive care unit.Twenty consecutive patients were included. The mean ARISCAT score was 41. The mean right diaphragmatic excursion was significantly increased with increasing inspiratory flow . Linear regression analysis showed a dependent speed increase without ceiling effect. Lung volumes were also significantly increased compared to baseline, with increasing total flow  and equally PaO2 . A significant decrease in respiratory rate was observed . No difference were noted in the modified score Borg and the right or left side.Application of progressive flow through HFNC improves diaphragmatic excursion, lung volumes and oxygenation while decreasing respiratory rate, speed-dependent manner. Its clinical impact remains to be seen in the population of high respiratory risk patients undergoing major abdominal surgery.


Stress- induced cardiomyopathy is acute, reversible left ventricle mainly apical dysfunction in patients with normal coronary angiography. Rarely it regards basal segments, therefore defined as “inverted stress- induced cardiomyopathy”. While classic form mostly affects postmenopausal women, inverted variant occurs essentially in younger females, always triggered by stress. It can also develop after medical procedures and surgery. Herein we report such unique case of 36- year old woman after cesarean delivery.A 36- year- old white woman at 40 week of gestation was admitted to hospital for elective repeated cesarean delivery. During caesarean delivery under spinal anaesthesia a previously healthy woman became hypotensive, requiring ephedrine to maintain her blood pressure. Three hours after delivery the patient presented acute heart failure and pulmonary oedema. Due to low blood pressure she demanded the administration of inotropic agents. Owing to respiratory failure and gradual deterioration of consciousness, mechanical ventilation was applied. Results of additional tests finally excluded pulmonary thromboembolism and acute coronary syndrome. The transthoracic echocardiography revealed severe left ventricular systolic dysfunction, ejection fraction 30 % with hypokinesis of the mid and basal segments of posterior, anterior and lateral wall with preserved contractility of the apical segments. The diagnosis of inverted stress- induced cardiomyopathy was set upon the overall clinical data. Both echocardiography and magnetic resonance imaging performed on the fifth day showed complete recovery of myocardial function. The patient was discharged after 15 days in good overall condition. At 12- month follow- up she remained asymptomatic with no echocardiographic abnormalities.Inverted stress- induced cardiomyopathy may occur in postpartum period, especially in combination with spinal anesthesia and adrenergic stimulants administration. The clinical awareness and multimodality imaging of possible diagnosis and further management of this unexpected variant of acute heart failure after caesarean delivery is required. Stress- induced cardiomyopathy (SIC) also known as “takotsubo cardiomyopathy” is characterized by reversible left ventricular dysfunction with chest symptoms (pain or dyspnea), electrocardiogram changes that mimic those of acute coronary syndrome (ACS) and minor elevation in serum levels of cardiac enzymes in patients with normal coronary angiography . While eWe report an extremely rare case of inverted SIC which occurred in young women after cesarean delivery.A 36- year- old white woman at 40 week of gestation was admitted to a local obstetric clinic for elective repeated cesarean delivery due to slanting position of the fetus. Both her previous and current pregnancy were uncomplicated. Her past medical history was unremarkable and no family history of cardiac disease was reported. The caesarean delivery was performed following the administration of standard spinal anesthesia. During surgery the patient became hypotensive and ephedrine was injected to maintain her blood pressure in the normal range. Three hours after delivery of healthy male infant, the patient complained of nausea, increasing dyspnoea and palpitations. On physical examination tachycardia with ventricular extra systoles and pulmonary rales were detected. Her blood pressure was 80/40 mmHg and demanded the administration of inotropic agents . Owing to worsening of oxygen saturation up to 70 % and gradual deterioration of consciousness, mechanical ventilation was applied and the patient was transferred to intensive care unit. Chest X ray indicated pulmonary congestion. Emergency computer tomography excluded pulmonary thromboembolism (PE) and confirmed severe pulmonary oedema Fig. . The patIt has been shown that SIC predominantly affects postmenopausal woman . AlthougThe pathophysiological mechanism of SIC is still not completely understood. Several possibilities have been discussed but increased levels of catecholamine and vasoconstrictive substances appear to play crucial role in triggering SIC . It can Minatoguchi et al. identified a total of 18 (8 from Western countries) confirmed cases of symptomatic P- SIC . Most woIn conclusion, this case shows that inverted SIC may occur in young women, particularly in postpartum period, especially in combination with spinal anaesthesia and adrenergic stimulants administration.On the basis of our experience we recommend the clinical awareness of possible diagnosis and further management of this unexpected variant of acute heart failure after caesarean delivery. Although initially the inverted stress- induced cardiomyopathy may seem dramatic, proper and accurate diagnosis usually leads to patients positive and successful outcome.I confirm that the study has been approved by Medical University of Lodz Ethics Committee.I confirm that formal written consent to publish all participant’s data was obtained from individuals involved in the study.


Comprehensive genomic and proteomic analyses demonstrate that there is nearly universal activation of the PI3K pathway in glioblastoma (GBM) patients . PersistOncogenic signaling pathways directly promote metabolic reprogramming to upregulate the biosynthesis of proteins, nucleotides, amino acids and lipids, required for the enhanced growth of cancer cells. These alterations include aerobic glycolysis known as the Warburg effect, which provides cancer cells selective advantages through enhanced catabolism of glutamine . GlutamiWe recently examined the role of glutamine metabolism in response to mTOR-targeted treatments for GBM . SurprisThese results demonstrate that the inhibition of mTOR signaling is sufficient to change the metabolic characteristics of GBM cells. Increasing GLS expression to elevate glutamate levels could be a method of compensating for the loss of TCA intermediates that occurs when mTOR inhibition reduces glycolysis. It is possible that mTOR inhibition may also induce the use of glutamine carbon sources to sustain the TCA cycle. Glutamate-derived αKG can be metabolized through the oxidative TCA cycle and/or reductively catabolized into isocitrate and citrate . It willAnother open question concerns the functional link between mTORC1 and glutamine metabolism. A recent study demonstrated that mTORC1 inhibition decreases glutamine metabolism by upregulating SIRT4 expression to suppress GDH activity . We founGLS has two isozymes in humans: kidney-type glutaminase (KGA) and liver-type glutaminase (LGA), which are encoded by GLS and GLS2 genes, respectively . GLS hasGBM is the most common adult malignant primary brain tumor and is one of the most lethal cancers. The median survival for GBM patients is 12-18 months from the time of initial diagnosis despite surgery, radiation and chemotherapy. Therefore, new therapeutic approaches are desperately needed. Recent genomic and proteomic analyses have developed biomarker-driven strategies and potential “GBM-specific molecular targets” have been applied over the last several decades. However, drug resistance to this category of therapeutics has led to minimal clinical efficacy to date. Our study provides new insight into GBM resistance mechanisms to targeted therapies and offers a compelling rationale for the simultaneous inhibition of mTOR and GLS as a promising combination therapy for this challenging brain cancer.


Medical flow cytometry (FACS) provides diagnostic answers by detecting the presence and concentration of cell populations, and/or by measuring concentrations of cell surface markers expressed on cells. Currently, FACS is limited to high complexity laboratories with time consuming pre-analytical steps, requiring trained technologists available only during business hours. The Accellix table top flow cytometer automates the process with results available within 20 minutes. Sample preparation and reading are performed in a dedicated disposable cartridge. Analytical data processing utilizing proprietary algorithms provides answers directly to the user.The Accellix disposable cartridge-based platform implements sample preparations using three reagent blisters. The three Accellix CD64 cartridge blisters contain staining cocktail of conjugated monoclonal antibodies, lysis buffer and reference beads respectively. Once sample processing is complete, the sample flows through a dedicated reading channel where data is acquired. The present study compared the results of neutrophil CD64 levels performed by standard laboratory FACS and the Accellix system in ICU infected and control patients and normal volunteers.In a demonstration of cell surface marker quantitation a comparison study of 118 blood samples showed a correlation coefficient of 0.94 for Accellix determined neutrophil CD64 compared to those determined using a FACS. The comparison of the CD64 levels performed by the FACS and the Accellix system are shown in figure


Such an open forum will help focus on novel annotations and on optimally benefiting from the energy of many experts. As proof-of-concept, we annotated protein subcellular localization in 100 abstracts cited by UniProtKB. The detailed comparison between our new corpus and the original UniProtKB annotations revealed sustained novel annotations for 42% of the entries (proteins). In a unified linked annotation resource these could immediately extend the utility of text corpora beyond the text-mining community. Our example motivates the central idea that linked annotations from text corpora can complement database annotations.Annotators of text corpora and biomedical databases carry out the same labor-intensive task to manually extract structured data from unstructured text. Tasks are needlessly repeated because text corpora are widely scattered. We envision that a The natural language processing (NLP) and biomedical research communities have in common that they invest great effort into making high-quality manual annotation of biomedical literature. The focus and the annotation strategies of the two communities have, however, differed so much that collaborations remained stunningly limited. Most text corpora contain detailed markup of only a few types of entities and relationships in a limited number of abstracts or articles   .model organisms: Homo sapiens (50 entries), Saccharomyces cerevisiae (baker's yeast with 25 entries), and Arabidopsis thaliana as a plant (25 entries). We used 46 of the 100 abstracts to develop our annotation guidelines that are available at https://www.tagtog.net/-corpora/loctext.We assembled a corpus of 100 PubMed abstracts referenced by UniProtKB. We focused on three tagtog system  [http://reflect.ws) [https://www.tagtog.net/-corpora/loctext under the Creative Commons Attribution 4.0 (CC-BY 4.0) license.Two of us (TG & SV) then annotated the remaining 54 abstracts. The two annotations agreed at F1 = 94% for entities and at F1 = 80% for relationships. We normalized protein names to UniProtKB and localizations to GO identifiers. The resulting corpus contains 306 annotated relationships in 201 different UniProtKB proteins with 48 GO distinct localization terms. All annotations were made within the framework of the Linked annotations from text corpora can complement database annotations only if manual corpus annotations identify relationships not captured by existing databases. Therefore, all our annotations were done from scratch without using database annotations. Comparing our "from scratch" annotations with those from UniProtKB revealed important novelty added by our text corpus.Arabidopsis RabF2a (UniProtKB entry RAF2A_ARATH) is localized to endosomes  proteins in 34 abstracts  compliant.We envision a linked annotation resource to continuously grow, supported by annotation tools making it easy for corpus developers to link future annotations; for example, through a standard JSON format. Not all linked annotations need to be made manually, though. Including also results from automatic text mining pipelines would help address the challenge of the prohibitively high costs of large-scale manual annotation . Associa


Focused ultrasound (FUS) can penetrate soft tissue noninvasively and induce physiological effects deep within target tissues. MR-image guidance, lesioning precision, sparing of healthy tissue and absence of ionizing radiation make FUS an ideal modality for brain interventions. Based on the encouraging results in treating patients with chronic pain or movement disorders through thermal ablation of thalamic and subthalamic targets, noninvasive transcranial MR guided focused ultrasound (tcMRgFUS) recently received CE marking for functional neurosurgery. Here, we report the world’s first successful application of noninvasive tcMRgFUS for tumor ablation in a patient suffering from a centrally located malignant glioma.Case Report: A 63 year old patient presented in our clinic with tumor recurrence in the left thalamic and subthalamic region five years after first surgery for a glioblastoma.® system  integrated into a clinical 3T MR system . The patient received local anesthesia for positioning a stereotactic frame , and prophylactic administration of paracetamol and ondansetron for preventing pain or nausea. Post-interventional assessment included follow-up MRI immediately after and on days 1, 5 and 21 after the procedure. The patient was awake and responsive during the whole intervention with stable neurological assessment and without additional medication needed. In the 3 hours lasting procedure, a total of 25 sonications were applied with up to 19’550 Joules of acoustic energy. 17 sonications reached ablative temperatures of over 55°C with a maximum of 65°C, as recorded by intraoperative realtime MR thermometry. MRI scans acquired immediately after the intervention revealed significant lesions to the sonicated tumor tissue that were particularly well visible in DWI and ADC maps. Later on, contrast-enhanced MRI showed well-circumscribed volumes of non-perfused tissue, indicative for ablated tumor tissue. As expected from earlier experience, perifocal edema developed around the thermal lesions, but gradually disappeared during the follow-up period. Neurological examination on day 5 revealed improvement of the preexisting right arm paresis while no additional neurological deficits were observed.After giving informed written consent he was included in our ongoing clinical phase-1 study on the feasibility and safety of tcMRgFUS for the treatment of brain tumors. Due to the location of the tumor within eloquent brain areas and considering previous radiotherapy and various cycles of different chemotherapeutics, surgical resection was excluded as a treatment option. The tcMRgFUS procedure was performed using the ExAblate NeuroThe successful ablation of a brain tumor demonstrates the feasibility of noninvasive tcMRgFUS tumor surgery. Further clinical interventions in the context of our ongoing clinical phase I study will be needed to assess the safety and efficacy of tcMRgFUS for brain tumor treatment and its relevance for future treatment strategies against brain tumors.