Prostate cancer (PCa) is the second most common cancer in men and an estimated 1.1 million men worldwide were diagnosed with prostate cancer in 2012 with almost 70% of the cases occurring in more developed regions [Random systematic prostate sampling with TRUS guided biopsy also has its inherent limitations. First of all, clinically insignificant cancers are often identified by chance and affect survival data due to lead and length time bias from overdetection and overtreatment of indolent disease . SecondlImaging has always been problematic in respect to PCa diagnosis and has been essentially limited to biplanar transrectal ultrasound and its modifications. Technological ultrasound developments with contrast enhancement and 3D reconfiguration have been underutilized by the urological community as a whole. Isotopic efforts with FDG and 11-choline PET scans have also disappointed in terms of diagnostic yield of prostatic disease. Recently, with the advent of better technology and different sequences, magnetic resonance imaging (MRI) has shown to be able to play a significant role in this respect. Numerous studies have now indicated that multiparametric (Mp) prostate MRI at 3 Tesla, including anatomical and functional sequences, enables accurate PCa detection and local staging with reasonable sensitivity and specificity . Given tSensitivity for detecting larger (>5 mm) and more aggressive (Gleason score > 7) tumors is better for Mp-MRI, indicating that it may preferentially detect clinically relevant tumors . HoweverIn an effort to standardize the reporting of Mp-MRI findings and to reduce subjectivity of image interpretation, the PI-RADS (prostate imaging and reporting archiving data system) classification has been developed. In this scoring system every parameter  is scored on a five-point scale. Additionally, each lesion is given an overall score, to predict its chance of being a clinically significant cancer . NeverthMp-MRI represents a potential tool to address the limitations of contemporary random systematic biopsy related to undersampling and false-negativity. Among biopsy-naive men, it increases the frequency of significant cancer detection to 50% in low-risk  and 71% in high-risk  patients .Among active surveillance candidates, repeat biopsy using MRI targeting demonstrates high sensitivity in confirming low-risk disease in low suspicion score lesions and risk upgrading in highly suspicious lesions .Techniques of MRI targeted biopsy include visual cognitive transrectal ultrasound guided biopsy, software coregistered magnetic resonance imaging-ultrasound, transrectal ultrasound guided biopsy, and in-bore magnetic resonance imaging guided biopsy. While many studies have compared targeted biopsy to systematic biopsy, few have evaluated the cancer detection rates across different targeted techniques. Fusion targeting seems to improve accuracy for smaller MRI detected lesions as well as high-grade cancer (Gleason score > 7) . HoweverMp-MRI is also a feasible and very useful tool in detecting local recurrence in patients who have undergone radical prostatectomy (RP) or radiotherapy. Concerning post-RP local recurrences, DCE-MRI seems to be the most reliable technique for detection, though DWI can be proposed as a reliable alternative . For preIs Mp-MRI ready for prime time? Well, this question remains open. Currently, the use of Mp-MRI as a “screening” tool for men referred with abnormal PSA for biopsy decision-making cannot be a general recommendation. However, it will undoubtedly have a place alongside novel biomarkers in diagnostic and surveillance strategies, in repeat biopsy settings, and perhaps in surgical planning for nonmetastatic bulky local disease as part of a planned multimodality treatment regimen.We hope that the readers of this special issue with 11 manuscripts which explores different aspects of Mp-MRI for diagnosing and treating PCa in its different forms will find some answers for different clinical settings. We are confident that this very timely compilation will be a major reference for many of us interested in prostate cancer.


While a rich body of research in controlled experiments has established changes in the neural circuitry of emotion in major depressive disorders, little is known as to how such alterations might translate into complex, naturalistic settings - namely involving dynamic multimodal stimuli with rich contexts, such as those provided by films. Neuroimaging paradigms employing dynamic natural stimuli alleviate the anxiety often associated with complex tasks and eschew the need for laboratory-style abstractions, hence providing an ecologically valid means of elucidating neural underpinnings of neuropsychiatric disorders. To probe the neurobiological signature of refined depression subtypes, we acquired functional neuroimaging data in patients with the melancholic subtype of major depressive disorder during free viewing of emotionally salient films. We found a marked disengagement of ventromedial prefrontal cortex during natural viewing of a film with negative emotional valence in patients with melancholia. This effect significantly correlated with depression severity. Such changes occurred on the background of diminished consistency of neural activity in visual and auditory sensory networks, as well as higher-order networks involved in emotion and attention, including bilateral intraparietal sulcus and right anterior insula. These findings may reflect a failure to re-allocate resources and diminished reactivity to external emotional stimuli in melancholia. As minimum training or in-scanner performance is required, this approach enjoys similar advantages to resting state acquisitions and thus holds great potential for imaging patient populations, minimizing anxiety associated with completing difficult or repetitive tasks. On the other hand, natural stimuli put ecologically relevant constraints on neuronal processes and are thus more likely to selectively engage brain networks of interest than resting state acquisitions. Here, we acquired fMRI data while patients with the melancholic subtype of MDD (henceforth melancholic-MDD) and age-matched healthy controls freely viewed film clips with different emotional salience . We chos2We analyzed fMRI data from 23 healthy and 17 participants with melancholia who viewed two emotionally salient film clips and one neutral film. The positive emotional film was an out-take from a stand-up comedy show by Bill Cosby, a well-validated stimulus shown to consistently elicit smiling and laughterFree viewing of these films evoked robust and consistent neural activity, as quantified by the inter-subject correlation (ISC) of voxel-wise BOLD signals , left. Rp < 0.05, FDR corrected). This reduction encompassed sensory association areas, including medial occipital gyrus (mOG) and superior temporal gyrus (STG), as well as high-order brain regions, including bilateral IPS, right anterior dIns and IFG. Intriguingly, the responses of vmPFC were significantly reduced in patients with melancholic-MDD specifically during free-viewing of the negative film , to extract large-scale network patterns that were engaged during natural viewing. Within each film condition, tICA identifies distributed brain regions whose neural activities co-vary across participants, without Several of these functional network components showed similar patterns to those identified in resting state fMRI data, confirmed through spatial cross-correlation with previously published templates and visual inspection2930The temporal profile of functional network activity revealed that distributed cortical regions were collectively engaged during selective aspects of complex natural scenes. One representative network of note is an auditory-visual network (component 10) derived from the negative film viewing condition. This network encompasses both auditory regions, including STG and Heschl’s gyri, and the ventral visual stream, including FUS . Based oObserving the activity of this sensory network motivated us to examine networks encompassing higher-order cortical regions. Component 5 from the negative film viewing condition had a spatial distribution consistent with the dorsal attention network 33. Its t1234p < 0.05, FDR corrected). There were no significant reductions during neutral film viewing. This pattern of group differences  converged with the results from voxel-wise ISC analyses, suggesting that melancholia is associated with substantial disturbances in the neuronal underpinnings of negative emotional experience.The dynamics of functional networks derived with tICA exhibited diminished inter-subject consistency in melancholic-MDD participants during the emotional films. In particular, we tested for group differences in ISC for the first 10 network components from each film viewing condition, controlling for multiple comparisons using false discovery rate (FDR). ISC was significantly reduced in melancholic-MDD for 8 out of the 10 network components during negative film viewing, and 3 network components during positive film viewing (p = 0.02). Crucially, within our depressed group, this variability was negatively correlated with QIDS scores, a standardized clinical instrument measuring depression severity . This correlation remains significant when age, gender, current smoking status, age at onset, duration of current episode and family history are included as covariates . We note that this (correlation) analysis is independent of the original whole brain search (which did not incorporate symptom severity in any way). BOLD signal variability in voxels outside of the ISC difference map did not differ significantly between melancholic-MDD and HC participants (p > 0.05), and were not correlated with depression severity . Such specificity suggests that the differences in neural activity were not due to physiological confounds or motion artifacts. Visualizing the distribution of the data points from patients with their various medications, shows a lack of clustering by medication class, suggesting that these results were unlikely to have been driven by specific medication effects  in MDD11 effects .By capturing the specific deficits in emotional circuit activity during free viewing of emotionally salient film clips, our findings bring fresh insights to the functional neurobiology of the melancholic sub-type of major depressive disorder. Our work recasts established findings using controlled stimuli into a more ecologically valid context. We observed a range of context-specific changes in sensory cortices, including FUS, PPA, mOG and STG, as well as higher order brain regions such as vmPFC, anterior insula and IPS – key regions in emotion processing and salience detection351213Naturalistic paradigms, such as films, offer enormous potential to reveal neural processes underlying dynamic emotional responses and their perturbation in neuropathological conditions. Such designs could be more effective in revealing brain dysfunction than resting state paradigms, which have been used extensively over the last ten years to study neuropsychiatric disorders574041424354041437The deployment of both voxel-wise and multivariate analyses using tensor ICA revealed convergent findings. Tensor ICA provides a novel method to extract large-scale functional networks driven by natural stimuli. In contrast to the use of spatial ICA in previous neuroimaging studies with natural stimuli18dwell – in non-adaptive states in melancholic individuals, and are unable to engage dynamically as they do in healthy control participants. The reduced variability in this key region also mirrors the subjective reports of those with this classic sub-set depressive condition, namely an impoverished cognitive experience, with persistent dysphoria that is unable to be shifted by external stimuli, either positive or negative in content – namely anhedonia. It is possible that the disturbances in perceptual regions in our results might be secondary, subject to impaired top-down control from vmPFC. Imaging analysis using effective connectivity could shed further insights on these possibilities.The reduced consistency in neural responses to the negative film in melancholic-MDD could reflect the altered internal processing of negative emotion – patients with depressive disorders, although they share a common diagnosis, are likely to process negative stimuli in distinctive ways that are unique to their personal experience, resulting in reduced consistency in neural responses to negative stimuli. Further, the robust correlation between the reduction in vmPFC dynamics during film viewing and the severity of the depressive episode suggests that these regions are stuck – or Patients with melancholia were on antidepressant and/or mood stabilizer and antipsychotic medications. It is thus possible that our findings are confounded by medication. Clinical research on melancholic depression is challenged by a number of barriers to control for this – the severity of the disorder prohibits withholding of medication for research purposes, whilst the variety of classes and pharmacological targets prevents the possibility to estimate a class-invariant dosage equivalent. We have visualized the distribution of data points from patients using various medications . The lacPhysiological responses are an integral part of emotional experiences4619In sum, using naturalistic, emotionally salient material, we highlight several key regions whose activities showed diminished consistency and variance in patients with melancholia. Such results - and the simple approach of showing movies to patients - has considerable clinical potential; assisting diagnosis, monitoring response to treatment and, possibly, even planning brain stimulation approaches in patients with treatment-resistant illness.The study was carried out in accordance with the approved protocol by the University of New South Wales ethics committee (HREC 08077), and all subjects provided written informed consent prior to participation.Functional neuroimaging data were acquired from forty participants  for this study. Seventeen patients  with major depressive disorder (MDD) were recruited through the specialist depression clinic at the Black Dog Institute in Sydney, Australia. Twenty-three right-handed age matched healthy controls  were recruited from the Sydney community, and who disavowed lifetime or current mood and/or psychotic illness. All participants were screened for mood and psychotic conditions with the Mini International Neuropsychiatric Interview (MINI). Clinical MDD participants comprised those meeting criteria for a current major depressive episode, but without lifetime (hypo)mania or psychosis being identified during the MINI interview. We restricted our analyses to patients diagnosed with melancholic depression, made by clinic psychiatrists using previously detailed criteria2Exclusion criteria for all participants were a current or past drug or alcohol dependence, ECT in the past six months, neurological disorder , brain injury , invasive neurosurgery, and/or an estimated full scale IQ (WAIS-III) score below 80 as assessed by the Wechsler Test of Adult Reading (WTAR).Depression severity in the clinical group was quantified with the Quick Inventory of Depressive Symptomatology or QIDS-SRFor the positive condition, participants viewed a stand-up comedy routine by Bill Cosby . For the negative condition, participants viewed a scene from the drama film “The Power of One”, which depicts the inhumane treatment of prisoners under the apartheid era. For the neutral condition, participants watched dynamic video footage of landscapes (recorded from a moving plane) and moving streams of water with matching audio streams. Participants watched the films through an MRI-compatible monitor, and were provided with brief instructions (displayed on the screen) prior to the onset of each movie (“Video will begin soon. Please relax and watch.”). High-quality audio from the movies was provided through insert earphones using an MRI-compatible system (Sensimetrics Model S14). All video stimuli are available from the authors on request.Structural and functional MR images of healthy controls and melancholic-MDD patients were acquired on a Philips 3 Tesla scanner equipped with a 12-channel receiver head coil. Functional images were pre-processed using the SPM8 toolbox29How to cite this article: Guo, C. C. et al. Out-of-sync: disrupted neural activity in emotional circuitry during film viewing in melancholic depression. Sci. Rep.5, 11605; doi: 10.1038/srep11605 (2015).


Trastuzumab is a monoclonal antibody highly effective in the treatment of several cancers, but its use is associated with cardiac toxicity which usually responds to cessation of the drug and/or medical therapy. We present an unusual case of acute cardiac toxicity temporally related to administration of trastuzumab in which the clinical presentation suggested an acute coronary syndrome. Coronary angiography, however, demonstrated minimal epicardial disease, but new wall motion abnormalities. Furthermore, the patient did not respond to withdrawal of the drug or medical therapy for heart failure. Trastuzumab is a monoclonal antibody part of standard medical therapy for both early and advanced breast cancer in individuals whose tumors overexpress the human epidermal growth factor receptor 2 (HER2) protein . HER2 isCardiotoxicity has been reported to occur with trastuzumab when administered alone and in combination with antineoplastic agents, particularly anthracyclines . The risWe report a case of trastuzumab-associated cardiotoxicity, initially manifesting as an acute coronary syndrome (ACS), in a 59-year-old male with metastatic adenocarcinoma of the esophagus. The patient was diagnosed in October 2010 (20 months prior to his ACS admission). His initial treatment involved cisplatin, 5-fluorouracil, and radiation followed by esophagogastrectomy and bilateral myotomy. Progression of his disease was noted the following year in April 2012. He was initiated on FOLFOX  and trastuzumab for HER2-positive disease. The FOLFOX was given every two weeks and the trastuzumab every three weeks.Immediately following his third trastuzumab infusion, he developed chest tightness, shortness of breath, and nausea. An EKG demonstrated new anterolateral T-wave inversions . SignifiThis case report is unique in that we describe a male undergoing chemotherapy with trastuzumab who presents with findings mimicking an acute coronary syndrome. Thus far, other similar case reports have described only women , 8. ThesThe pathogenesis of trastuzumab-associated cardiotoxicity is not completely understood. Unlike anthracycline cardiotoxicity where cardiac damage is believed to result from free radical injury and oxidative stress resulting in myocyte loss, trastuzumab cardiotoxicity is believed to result from blocked ErbB2 signaling pathways. The proto-oncogene ErbB2  encodes a receptor tyrosine kinase critical in cardiovascular development. Genetically manipulated mice lacking ErbB2 die in utero with failure of cardiac development. In the postnatal heart, ErbB2 conditional knockout mice develop dilated cardiomyopathy at eight weeks of life. How this occurs at the molecular level remains under active investigation. Expression of ErbB receptors decreases in chronic heart failure in humans, suggesting that altered ErbB signaling plays a role in the progression of heart failure .Important lessons emerge from this unique case report. First, we describe a case of trastuzumab cardiotoxicity mimicking ACS presenting in a man, which is different from those previously described in women. Numerous studies have reported a difference in heart failure between men and women, thus suggesting that variable expression of myocyte receptors (or ligands) in men and women could be involved in the development and progression of heart failure. Unlike other studies of trastuzumab cardiotoxicity, this patient appears to have had irreversible cardiac damage despite minimal epicardial disease, standard heart failure treatment, and cessation of the drug. Clinicians should be aware of this possibility when caring for patients with trastuzumab-associated cardiotoxicity.


Posttraumatic Stress Disorder (PTSD) is characterized by intrusive recall of the traumatic memory. While numerous studies have investigated the neural processing mechanisms engaged during trauma memory recall in PTSD, these analyses have only focused on group-level contrasts that reveal little about the predictive validity of the identified brain regions. By contrast, a multivariate pattern analysis (MVPA) approach towards identifying the neural mechanisms engaged during trauma memory recall would entail testing whether a multivariate set of brain regions is reliably predictive of  whether an individual is engaging in trauma or non-trauma memory recall. Here, we use a MVPA approach to test 1) whether trauma memory vs neutral memory recall can be predicted reliably using a multivariate set of brain regions among women with PTSD related to assaultive violence exposure (N=16), 2) the methodological parameters  that optimize classification accuracy and reproducibility of the feature weight spatial maps, and 3) the correspondence between brain regions that discriminate trauma memory recall and the brain regions predicted by neurocircuitry models of PTSD. Cross-validation classification accuracy was significantly above chance for all methodological permutations tested; mean accuracy across participants was 76% for the methodological parameters selected as optimal for both efficiency and accuracy. Classification accuracy was significantly better for a voxel-wise approach relative to voxels within restricted regions-of-interest (ROIs); classification accuracy did not differ when using PTSD-related ROIs compared to randomly generated ROIs. ROI-based analyses suggested the reliable involvement of the left hippocampus in discriminating memory recall across participants and that the contribution of the left amygdala to the decision function was dependent upon PTSD symptom severity. These results have methodological implications for real-time fMRI neurofeedback of the trauma memory in PTSD and conceptual implications for neurocircuitry models of PTSD that attempt to explain core neural processing mechanisms mediating PTSD. Posttraumatic Stress Disorder (PTSD) is characterized by re-experiencing of the traumatic event, avoidance of trauma-related stimuli, general changes in mood and cognition, and hyperarousal symptoms. PTSD haMeta-analyses that explicitly test the degree to which univariate functional activity within these regions is altered in PTSD provide partial support for these neurocircuitry models of PTSD. On the one hand, a recent fMRI meta-analysis  focusingOne approach towards clarifying the neurocircuitry mediating PTSD is multivariate pattern analysis (MVPA) –14. In tA related field that would be informed by clearer delineation of core neural mechanisms mediating observable PTSD clinical symptoms is real-time fMRI (rtfMRI) neurofeedback ,17–20. IHere, we utilize a previously published data set  to conduParticipants consisted of 17 adult women with PTSD related to either physical or sexual assault. One woman moved excessively during the scan causing intractable signal artifact, and her data were subsequently removed from analyses. This resulted in a final sample of 16 participants. Inclusion criteria consisted of 1) a history of either physical or sexual assault, 2) a current diagnosis of PTSD, and 3) that participants were stable on any psychiatric medications for at least 4 weeks. Exclusion criteria consisted of psychotic disorders, a primary substance use disorder, or internal ferromagnetic objects. Participants were recruited from outpatient mental health clinics and from community wide advertisements. The assault exposure histories of participants were characterized using the trauma assessment section of the National Women’s Survey and National Survey of Adolescents –25. ThisPTSD and comorbid psychological disorders were assessed with the Structured Clinical Interview for DSM-IV Disorders (SCID)  administThis task was delivered in the context of study probing the neural correlates of repeated exposure to the trauma memory . Particip = .397).Participants were asked to indicate their level of anxiety, vividness of the memory, and dissociation during the memory recall on Likert scales of 1–10, and practiced using this rating scale prior to fMRI. The imagery procedure during fMRI consisted of five sequential presentations of the memory. The narratives were presented both aurally through headphones and visually, such that the written narrative was presented to them on a projected screen that participants viewed through a mirror attached to the MRI head coil. Each three minute presentation was preceded by 30 seconds of a resting-state, and these 30 seconds were not used in the classification analyses described below. Participants provided the subjective ratings after each individual presentation of each narrative. The trauma and neutral narratives were presented sequentially , with the order  counterbalanced across participants. The two script types were separated by a 4 minute anatomical scan. Preliminary analyses demonstrated that anxiety during the first neutral script was not significantly higher among women who received the trauma scripts first compared to the women who received the neutral scripts first (3 resolution). Echo planar imaging (EPI) sequences were used to collect the functional images using the following sequence parameters: TR/TE/FA = 2000ms/30ms/90°, FOV = 240x240mm, matrix = 80x80, 37 oblique slices , slice thickness = 3 mm, final resolution = 3x3x3 mm3. Each memory presentation was collected as a separate EPI run .A Philips 3T Achieva X-series MRI system using an 8-channel head coil  was used to acquire imaging data. Anatomic images were acquired with a MPRAGE sequence , CSF, and WM were segmented from the high resolution anatomical image using FSLImage preprocessing followed standard steps and was completed using AFNI  softwareC parameter = 100. This algorithm attempts to find a decision boundary differentiating the classes of stimuli  based on the nonlinear combination of features. SVM was implemented in Matlab using the LIBSVM library [Across all methodological permutations tested, we used a common cross-validation procedure and classification algorithm. We used support vector machines (SVM) to train a classifier to discriminate trauma memory recall from neutral memory recall using the voxel-wise fMRI timeseries as the input features. The SVM algorithm used a non-linear radial basis function kernel and  library . The feaWe tested classification accuracy across the following permutations: 1) spatial smoothing vs no spatial smoothing, 2) number of memory recall repetitions (range 1–5), and 3) brain mask restricting which voxels are used in the SVM classifier .Among datasets that were not spatially smoothed, the last step of the preprocessing pipeline was detrending, whereas among spatially smoothed data sets an additional spatial smoothing step  was included.All participants underwent 5 successive memory recall repetitions for each script type. We tested classification accuracy and reproducibility of the SVM feature weight spatial maps between models trained using the first run of each script type, the first two consecutive runs, the first three consecutive runs, the first four consecutive runs, and all five runs.We compared classification accuracy between models trained using all GM voxels, only voxels within ROIs specified in PTSD neurocircuitry models, all GM voxels except those voxels within ROIs specified in PTSD neurocircuitry models, and only voxels within randomly generated ROIs. The GM mask was defined by segmenting each subject’s high resolution T1 image (using FSL) into GM, white matter, and CSF, and then selecting those GM voxels that are shared by at least half of the participants. This resulted in a common GM mask used across participants consisting of 36280 voxels.As indicated in the introduction, all current neurocircuitry models of PTSD implicate the amygdala, hippocampus, and medial PFC as key neural mechanisms mediating PTSD, and a recent meta-analysis additionally implicated the retrosplenial cortex, PCC, and precuneus in PTSD re-experiencing symptoms specifically. The specific anatomical coordinates of the PTSD ROIs used here were derived empirically from two recent meta-analysis of functional activation studies in PTSD ,9: left To foster comparisons regarding the explanatory power of the PTSD ROIs, we also compared classification accuracy to two control masks: 1) a mask using all GM voxels except for those implicated in PTSD neurocircuitry models , and 2) voxels within 8 randomly generated ROIs  within the GM mask . Regarding the randomly generated ROIs with the GM mask, we generated 10 sets of 8 randomly generated ROIs , and tested classification accuracy for each participant using each of the 10 sets of random ROIs, and then defined classification accuracy for the random ROIs as the median cross-validation accuracy across all 10 random sets of ROIs.Spatial maps representing each voxel’s contribution to the nonlinear SVM decision function were created for each participant across each relevant methodological permutation by reshaping the SVM feature weights back into 3-D brain shape. Reproducibility of the maps across methodological permutation was assessed by examining the spatial correlation  between feature weights reshaped as 1-D vectors for a given participant. For example, for a given participant, we characterized the correlation of the SVM feature weights for the model trained using only the first run to the model trained using the first two runs.We also sought to assess whether there were commonalities in the feature weights contributing to the nonlinear SVM decision function across participants. This was assessed in two ways. First, we assessed univariate similarity for each voxel within the GM mask across participants using traditional mass univariate one-sample t-tests. We similarly used mass univariate robust regression analyses to test whether SVM feature weights scaled linearly with PTSD symptom severity (from the PCL) when controlling for age, comorbid depression, and comorbid substance use disorders. We corrected for multiple comparisons using cluster-level thresholding, with a corrected p < .05 achieved through a minimum of 47 contiguous voxels surviving an uncorrected p < .01 (based on 3dClustSim). Second, to maximize statistical power, we conducted parallel one-sample t-tests and robust regression analyses with PTSD symptom severity when constraining analyses within the 8 PTSD-related ROIs described above. The mean SVM feature weight among the voxels within these ROIs was calculated for each participant and then used in subsequent one-sample t-tests or robust regression analyses.Finally, we also conducted a control classification analysis to demonstrate that classification results and feature weight maps for differentiating trauma vs neutral memory recall cannot be explained by differences in motion-related artifact. In this analysis, we used framewise displacement (FD) to define motion across the 6 directions of motion displacement. FD refers to the sum of the absolute value of temporal differences across the 6 motion parameters; for example, an FD of .5 indicates a TR where the participant moved, in total across the 6 parameters, 5 mm. We used a median split to divide the FD into high motion TRs and low motion TRs, which were then the labels used in classification. The SVM classification analyses used the first three runs to parallel the analyses that seemed most efficient and accurate (see ps < .05). Using four runs and five runs was not significantly better than three runs. As depicted in r = .65); by contrast, similarity between SVM feature weights for the model trained with the first run and the model trained with all five runs for this same participant was less robust (r = .29). When this approach is repeated across all participants (We then assessed how similar (using r-to-z transformed correlation coefficients) the SVM feature weights were for a given participant across the models using different numbers of memory recall repetitions. icipants , SVM feaCombined these results suggest that the three and four run models had the greatest classification accuracy and yielded SVM feature weights with the greatest reproducibility. Given that repeated presentation of the trauma memory yields non-stationary brain dynamics, it seems that efficiency, reproducibility, and accuracy is best for the three run model. Thus, subsequent analyses are constrained to just models trained with three memory recall repetitions.t = .82, p = .42), suggesting that the classifier was not biased towards decisions of trauma or neutral memory recall.Beyond classification accuracy (hit rate), we also assessed sensitivity (proportion of trauma narrative TRs correctly identified as trauma narrative TRs) and specificity  of the three run model. Mean specificity across the 16 participants was 76%; mean sensitivity was 74%. There was no significant difference between sensitivity and specificity .As indicated in The purpose of the present study was to test 1) whether a multivariate set of brain regions could reliably discriminate between trauma and neutral memory recall among women with PTSD, 2) the methodological parameters that affect this classification accuracy, and 3) the correspondence between the brain regions that differentiate trauma recall and the brain regions predicted by PTSD neurocircuitry models. In regards to the first goal, we found that trauma memory recall could be differentiated reliably from neutral memory recall across women with PTSD based only on a snapshot (a single TR) of brain function. Mean classification accuracy across participants ranged from ~69%-77% for spatially smoothed data sets and were all significantly greater than chance. These basic results demonstrating significantly reliable classification accuracy have general implications. These data demonstrate that it is possible, and statistically reliable, to map a given individual with PTSD’s brain network encoding trauma memory recall. In a traditional univariate analyses, one would assess the statistical relationship between the task  and the activity in a voxel, and this is repeated across all voxels. While this approach identifies the effect of the task on individual voxel activity, it does not provide information regarding 1) co-activity across spatially distributed voxels , nor 2) the predictive validity of a voxel’s or set of voxels activity in differentiating the task. By contrast, these present results demonstrate both the spatially distributed patterns of voxel co-activity related to trauma memory recall and also the predictive validity of this network in differentiating trauma from neutral memory recall. Accordingly, the current study provides proof-of-concept evidence for applications of real-time fMRI neurofeedback to identify, and then target for volitional control, a brain-wide network encoding the trauma memory identified specifically for each individual.In regards to the methodological parameters that optimize classification accuracy, we observed that spatial smoothing had a marked advantage over non-spatial smoothing. This is counterintuitive in the sense that most MVPA analyses use nonsmoothed data sets and a putative strength of MVPA is utilizing spatial information that is lost during spatial smoothing –41. NoneWe also observed that using data from multiple runs was superior to using a single run, and that there appeared to be diminishing returns of increasing the number of runs after three runs. Beyond the observation that more data is clearly helpful in training the classifier to identify and separate the signal of interest, it is informative that increasing the number of runs beyond three does not improve classification performance. As noted above, the previous study from which these data come  demonstrWe also investigated the correspondence between brain regions differentiating trauma from neutral memory recall and the brain regions predicted by PTSD neurocircuitry models. We found that a whole-brain approach  was superior to an ROI approach. Further, while we found better than chance classification accuracy when using only voxels within PTSD neurocircuitiry-related regions, we also found that classification using PTSD neurocircuitry-related regions was no better than randomly generated ROIs, and that removing voxels with PTSD neurocircuitry-related ROIs did not weaken performance when using all other GM voxels. These results provide strong evidence that the brain networks encoding recall of the trauma memory cannot be reduced to regions canonically implicated in PTSD; rather, these data emphasize that distributed and brain-wide information processing mediates trauma memory recall. The GM voxel-wise group-level analyses failed to identify significant clusters of voxels where the SVM feature weights were similarly strong across individuals; however, the ROI analyses focused on PTSD neurocircuitiry-related ROIs demonstrated that the feature weights for the left hippocampus significantly differed from zero, and that SVM feature weights for the left amygdalohippocampal ROI were significantly negatively correlated with PTSD symptom severity. It is widely known that interpretation of SVM feature weights when using a non-linear kernel is difficult ,44. NoneThe current results have implications for the use of rtfMRI neurofeedback in the treatment of PTSD. Based on prior meta-analyses of either generic cognitive-emotional tasks or symptom provocation specifically ,9, one mThe present results demonstrate the feasibility of using MVPA to identify whole-brain networks that differentiate trauma from neutral memory recall among women with PTSD. We also demonstrate the methodological parameters that seem to optimize classification accuracy, with spatial smoothing and repeated memory presentations (with diminishing returns after a certain point) emerging as important factors affecting accuracy. We also found that whole-brain networks, rather than PTSD-specific ROIs, led to optimal classification accuracy, which emphasize the distributed nature of trauma memory recall and highlight that re-experiencing symptoms cannot be reduced to a few ROIs. These results also have practical implications for application of rtfMRI neurofeedback in PTSD. Nonetheless, the current study is not without limitation. First, this was an offline MVPA analysis, and thus any speculation regarding neurofeedback efficacy, with respect to either modulating a whole-brain vs ROI target or modulating clinical symptoms, is purely speculative. Second, the sample was limited to a small sample of women with assault-PTSD. The focus on women with assault-related PTSD makes the degree to which these results generalize to other samples of PTSD  is unknown, and the small sample size accordingly tempers inferences and calls for additional research to replicate and extend the current findings. Similarly, the lack of a trauma-exposed group without PTSD precludes any inferences regarding the degree to which the identified brain regions are unique to PTSD vs common to trauma-exposure per se cannot be made. While this is a limitation in making inferences regarding whether the neurocircuitry in PTSD differs from trauma-exposed without PTSD, note that this is not a limitation regarding the application of MVPA to the problem of differentiating trauma memory recall in PTSD. In light of these limitations, future work along these lines is needed to further probe the brain networks that mediating trauma memory recall in PTSD, the unique differences in these networks among PTSD samples (vs trauma-exposed controls), the methodological factors that optimize identification of the brain networks, and the efficacy of rtfMRI-based neurofeedback in modulating these networks and downstream clinical symptoms.S1 Fig(TIF)Click here for additional data file.


Brains achieve efficient function through implementing a division of labor, in which different neurons serve distinct computational roles. One striking way in which neuron types differ is in their electrophysiology properties. These properties arise through expression of combinations of ion channels that collectively define the computations that a neuron performs on its inputs and its role within its larger circuit. Though the electrophysiology of many neuron types has been previously characterized, these data exist across thousands of journal articles, making cross-study neuron-to-neuron comparisons difficult.Here, we present NeuroElectro, a public database where physiological properties for the majority of mammalian neuron types have been compiled through semi-automated literature text-mining and expert curation. The corresponding web application, at http://www.neuroelectro.org, provides a rich dynamic interface for visualizing and comparing physiological information across neuron types; conveniently linking extracted data back to its primary reference. Mining the database content after normalization for methodological differences, we show that there exist but 5-9 major neuron classes in terms of electrophysiological properties, which separate largely based on cell size and basal levels of excitability Figure .As an example of how this resource can help answer fundamental questions in neuroscience, we integrate NeuroElectro with neuronal gene expression from public datasets like the Allen Brain Atlas. We show that simple statistical models can accurately predict features of a neuron's electrophysiological phenotype given information of its gene expression alone. We further investigate these models to ask which genes, of the 20K in the genome, are most predictive of neuron physiology. We find that while ion channel-related genes provide significant predictive power, the most predictive gene classes surprisingly correspond to G-proteins and transcription factors, suggesting the involvement of hundreds of diverse genes in regulating a neuron's computational function.


Cancer is a multifactorial disease and is one of the leading causes of death worldwide. The contributing factors include specific genetic background, chronic exposure to various environmental stresses and improper diet. All these risk factors lead to the accumulation of molecular changes or mutations in some important proteins in cells which contributes to the initiation of carcinogenesis. Chemotherapy is an effective treatment against cancer but undesirable chemotherapy reactions and the development of resistance to drugs which results in multi-drug resistance (MDR) are the major obstacles in cancer chemotherapy. Strategies which are in practice with limited success include alternative formulations e.g., liposomes, resistance modulation e.g., PSC833, antidotes/toxicity modifiers e.g., ICRF-187 and gene therapy. Targeted therapy is gaining importance due to its specificity towards cancer cells while sparing toxicity to off-target cells. The scope of this review involves the various strategies involved in targeted therapy like-monoclonal antibodies, prodrug, small molecule inhibitors and nano-particulate antibody conjugates. Cancer is the second leading cause of deaths all over the world. Globally 7.6 million deaths are caused by cancer which represents 13% of all global deaths . SeveralTargeted therapy aims at delivering drugs to particular genes or proteins that are specific to cancer cells or the tissue environment that promotes cancer growth. Effectiveness of the therapy lies in targeted release of therapeutics at the disease site while minimizing the off-target side effects caused to normal tissues. It is often used in conjunction with chemotherapy and other cancer treatments. Targeted therapy involves developing drugs that block cancer cell proliferation, promote cell cycle regulation or induce apoptosis or autophagy and targeted delivery of toxic substances specifically to cancer cells to destroy them. Targeted therapy involves the use of monoclonal antibodies or oral small drugs .®; Wyeth, CT, USA), a CD-33 specific monoclonal antibody conjugated to a calicheamicin used for the treatment of acute myeloid leukemia [90Y metal isotope based anti-CD20 ibritumomabtiuxetan  has been developed for use in clinical therapy [Monoclonal antibodies are the focus of intense research in the field of cancer therapeutics since mid 1970s when the customized monoclonal antibody production was reported. Monoclonal antibody production, antibody engineering with display and screening innovations such as phage display meant the binding of antibody to a wide range of targeted antigens with exceptional specificity. Cancer immunotherapy involves the use of gemtuzumab  in tumor tissues by exogenous enzyme(s) which can be accomplished by several methods which include: gene-directed enzyme prodrug therapy (GDEPT), virus-directed enzyme prodrugtherapy (VDEPT), and antibody-directed enzyme prodrug therapy (ADEPT). The important aspect of prodrug cancer therapy is to deliver drug-activating enzyme or gene or functional protein to tumor tissues, followed by systemic administration of a prodrug .Prodrug cancer therapy is a two step process, the first step involves, targeting the drug-activating enzyme and its expression in tumors followed by the systemic administration of the nontoxic prodrug, which is the substrate for the exogenous enzyme that is targeted and expressed in tumors -18. ThisEscherichia coli CD/5-FC and E. coli NTR/CB1954 which act intracellularly by converting prodrugsinto active drugs within cancer. Cell-cell contact is essential for this mode of action for effective killing. An extra-cellular cytotoxic effector system includes the conversion of an inactive glucuronidated derivative of doxorubicin (HMR 1826) to the cytotoxic doxorubicin in the tumor cells by the secreted form of lysosomal human glucuronidase. In the extracellular system the hydrophilic prodrug gets converted into a lipophilic, cell-permeable cytotoxic drug outside cells and hence targets both transduced and nontransduced cells. It exhibits enhanced cytotoxic potential as cell-cell contact is not required for a bystander effect [Gene directed enzyme prodrug therapy, is a technique that involves delivery of a gene that encodes a foreign enzyme to tumor cells where it finds expression and activates a systemically administered nontoxic prodrug , 23, 24.r effect .E. coli NTR gene [Virus directed enzyme prodrug therapy (VDEPT) uses viral vectors to deliver a gene that encodes an enzyme that can convert a systemically administrated nontoxic prodrug into a cytotoxic agent within tumor cells. The NTR/CB1954 combination is used against colorectal and pancreatic cancer cells to sensitize them to CB1954 after retro-viral transduction and expression of the NTR gene , 26.in vitro or in vivo by VDEPT, the majority using CD/5-FC or HSV-TK/GCV with the involvement of retroviral and adenoviral vectors [The viruses used for VDEPT include: retroviruses, adenoviruses, HSV , adeno-a vectors .Genetic prodrug activation therapy (GPAT) induces the selective expression of a drug-metabolizing enzyme for activation of prodrug into a toxic moiety using the known transcriptional differences between normal and tumor cells , 34. SevAntibody directed enzyme prodrug therapy (ADEPT) uses a conjugate which consists of tumor specific antibody linked to a drug-activating enzyme which when administered systemically targets tumor tissues. This targeted enzyme which is localized on the tumor surface, converts the systemically administered nontoxic prodrug into a toxic drug resulting in cytotoxic effects in tumor cells , 35-40. The important criteria for ADEPT include: the target antigen should be accessible, therefore it should preferably be a membrane bound antigen associated with the tumor cell membrane or secreted into the extracellular matrix of the tumor , and theThe interval between enzyme and prodrug administrations is important for ADEPT, studies carried out in animals regarding the optimal interval showed that with the enzyme CPG2 linked to the anti-CEA antibody A5B7, the prodrug CMDA can be safely given 48 h or 72 h after antibody-enzyme administration . In humaThe limitations of ADEPT include: restricted delivery of the large conjugate in poorly vascularized tumors, therefore it is not possible to deliver antibody/enzyme conjugate to all of the tumor cells . With loRARα retinoic acid receptor gene with all-trans retinoic acid [BCR-ABL translocation with the ABL inhibitor imatinib [ALK gene [BRAF kinase [Small drugs constitute a pill that a patient takes orally. As they are smaller chemical components than monoclonal antibodies, the body absorbs them better. The small drugs usually are targeted against specific molecular targets which are important for cancer cell proliferation or metastasis or angiogenesis. The current state of cancer drug discovery and development focuses on small molecule inhibitors which act against new molecular targets that determine therapeutic outcome. The molecularly targeted cancer therapies have resulted in improving the lives of a large number of cancer patients. The successful treatment of patients with acute promyelocytic leukaemia harbouring translocations in the oic acid  and chroimatinib , 46 servimatinib . The recimatinib , crizotiALK gene  and vemuF kinase  for metaF kinase , 52. TheF kinase , 54.Despite the considerable progress in cancer therapy with the advent of new molecularly targeted therapies, therapeutic options are still limited for many patients and the process of new drug development is frustratingly slow with high failure rates , 55, TheAnother important task is identification of specific molecular targets through the sequencing of various cancer genomes which revealed extraordinary complexity with several genetic alterations and considerable genetic heterogeneity, not only between different tumours but also within an individual cancer -59. FurtAfter identification of a potential novel therapeutic target, the next challenge is that of selecting and validating the best targets. This requires establishing a causal linkage of the proposed target and target modulation to deliver a therapeutically meaningful biological effect in relevant experimental models. ‘Druggability gap’ is the main concern in the drug discovery for medicinal chemists using small molecules. Frequently, the promising targets are regarded as technically undruggable as they cannot be targeted with small molecules which is referred to as ‘druggability gap’ e.g., RAS proteins, c-MYC or hypoxia inducible factor (HIF) . UnfortuRecently research in nanoparticle based targeted therapy has gained momentum which saw the use of a full spectrum of nanoparticles in diagnostic and therapeutic applications of cancer . AntibodThus, the cancer drug discovery involves genome wide sequencing, understanding molecular pathology through bioinformatics and systems biology approaches for understanding how cancer cells can be targeted through single agents or on several fronts through drug combinations -75. Alth


The purpose was to assess if variation in sagittal plane landing kinematics is associated with variation in neuromuscular activation patterns of the quadriceps-hamstrings muscle groups during drop vertical jumps (DVJ).Fifty female athletes performed three DVJ. The relationship between peak knee and hip flexion angles and the amplitude of four EMG vectors was investigated with trajectory-level canonical correlation analyses over the entire time period of the landing phase. EMG vectors consisted of the {vastus medialis(VM),vastus lateralis(VL)}, {vastus medialis(VM),hamstring medialis(HM)}, {hamstring medialis(HM),hamstring lateralis(HL)} and the {vastus lateralis(VL),hamstring lateralis(HL)}. To estimate the contribution of each individual muscle, linear regressions were also conducted using one-dimensional statistical parametric mapping.The peak knee flexion angle was significantly positively associated with the amplitudes of the {VM,HM} and {HM,HL} during the preparatory and initial contact phase and with the {VL,HL} vector during the peak loading phase (p<0.05). Small peak knee flexion angles were significantly associated with higher HM amplitudes during the preparatory and initial contact phase (p<0.001). The amplitudes of the {VM,VL} and {VL,HL} were significantly positively associated with the peak hip flexion angle during the peak loading phase (p<0.05). Small peak hip flexion angles were significantly associated with higher VL amplitudes during the peak loading phase (p = 0.001). Higher external knee abduction and flexion moments were found in participants landing with less flexed knee and hip joints (p<0.001).This study demonstrated clear associations between neuromuscular activation patterns and landing kinematics in the sagittal plane during specific parts of the landing. These findings have indicated that an erect landing pattern, characterized by less hip and knee flexion, was significantly associated with an increased medial and posterior neuromuscular activation  during the preparatory and initial contact phase and an increased lateral neuromuscular activation  during the peak loading phase. Anterior cruciate ligament (ACL) injuries are very common during dynamic sports activities in the active population 16–39 years) accounting for approximately 26% of all internal knee injuries 9 years a. Non-conMany biomechanical risk factors for ACL injury have been proposed, however few studies have examined muscular activation patterns that might be related to ACL injury risk. Besides the external forces acting on the knee joint during dynamic activities, the quadriceps and hamstrings muscle groups have the potential to either load or unload the knee ligaments based on their coordinated activation. Several cadaveric  and in-vA prospective study by Zebis et al.  showed tDespite literature that suggested high risk neuromuscular activation patterns during DVJ, the relation between sagittal plane landing kinematics and muscular activation patterns of the quadriceps and hamstrings muscle groups is still not well examined. Recent work of Blackburn et al.  showed tIn the afore mentioned studies , 22, EMGTo date, only the relationship between individual muscle activations and landing kinematics has been investigated. However, as agonistic and antagonistic muscle pairs constantly interact, significant associations between neuromuscular activation and knee/hip joint flexion angles could be missed if only individual muscle activation is assessed. Therefore, we will use Statistical Parametric Mapping (SPM) in the present study, a method that accounts for inter-muscle covariance by creating anatomically relevant muscle groupings.We hypothesized that the variation in sagittal plane knee and hip landing kinematics is associated with the EMG activity patterns of the quadriceps and hamstrings muscle group. Based on the results of Blackburn et al.  and WalsFifty female athletes  consented to participate in this study . All participants were member of a Belgian elite level team  and were injury and pain free. Before participating in this study, all participants provided their written informed consent, which was approved by the local ethics committee. Thirteen participants were aged between 16 and 18 years and signed the informed consent themselves under the written permission of their parents, in which the parents indicated that they fully understood the content of the informed consent and agreed to the signature of their children. As such, no informed written consent was obtained from the next of kin, caretakers, or guardians on behalf of the minors/children enrolled in this study. This study conformed to the principles of the declaration Helsinki (1964), was approved by the local ethics committee and registered with reference number S53369. The local ethical committee approved the consent procedure used in this study. Additionally, all data regarding the participants were anonymized.Each test session started with a standardized warm-up, which consisted of two series of eight bipedal squats and eight bipedal jumps . ParticiDVJ are commonly used for screening in clinical settings to assess injury risk , 27,28. Each participant had 44 spherical reflective markers positioned according to the 6-degrees-of-freedom, eight segment ‘Liverpool John Moores University’ model (LJMU model) including feet, upper and lower legs, pelvis and trunk . SegmentA wireless EMG system  was used to record the muscle activity of the vastus lateralis (VL), vastus medialis (VM), hamstring lateralis (HL) (i.e. biceps femoris) and hamstring medialis (HM) (i.e. semitendinosus) using surface electrodes which were positioned according to the SENIAM guidelines. All electrode locations were shaved and gently cleaned with 70% isopropyl alcohol to reduce skin impedance. Silver-silver chloride, pre-gelled bipolar surface EMG electrodes  were placed over the muscle belly and aligned with the longitudinal axis of the muscle, with a center-to-center distance of 0.02 m. The minimum distance between electrode pairs was set at 0.03 m to reduce the possibility of cross-talk.2 AMTI  force plates. Force plate and EMG data were sampled at 1000 Hz. Three-dimensional kinematic data were simultaneously (time synchronized) recorded with the force and EMG data in Nexus  using 6 MX-T20 optoelectronic cameras  sampling at 100 Hz.DVJ were completed on two individual 0.8 x 0.3 mth order low pass Butterworth filter with a cut-off frequency of 18 Hz [th order zero-lag Butterworth filter at a cut-off frequency of 6 Hz. The EMG signal amplitudes of the DVJ Trials were normalized to the root mean square amplitude (over a period of 100 ms) of the MVC out of 3 attempts. Kinetic and kinematic data were normalized to 100% of the phase starting at 100ms before initial contact until take off as can be seen in Only the first landing (first contact) within each DVJ trial was used for analysis . Whole-bof 18 Hz . InitialPeak knee and hip flexion angles were calculated because these discrete values illustrate the amount of knee and hip flexion during the landing phase and are easy to measure in a clinical setting . The extAll participants (n = 50) were divided into quintiles based on their landing pattern. Quintile one consisted of the ten participants who demonstrated the highest peak knee/hip flexion angles. Quintile five consisted of the ten participants who demonstrated the lowest knee/hip flexion angles (more extended knee/hip joint angles). Quintiles 2, 3, and 4 demonstrated values ranging from much flexion towards less flexion, respectively.Shapiro-Wilk analyses were used to test normality of all kinematic and kinetic data of the different quintiles. Independent t-tests were used to compare the kinematics and kinetics between the upper and lower quintiles. Pearson correlation analysis was used to investigate the relation between peak hip and peak knee flexion joint angles in all participants.As we were interested in neuromuscular activation of muscle pairs around the knee rather than the activation of individual muscles, we created four anatomically meaningful EMG vectors: an anterior EMG vector field {VM,VL}, a lateral EMG vector field {VL,HL}, a posterior EMG vector field {HM,HL}, and a medial EMG vector field {HM,VM} as can be seen in 2 statistic.Furthermore, correlation analyses were used to further assess the relationships between peak knee/hip joint flexion angles and the neuromuscular activation patterns of VL, VM, HL and HM. As we did not want to reduce the EMG data to a discrete value, we used Statistical Parametric Mapping (SPM) a technique that allows us to analyze the entire EMG time series. To avoid multiple EMG signal co-variance bias ,24 we usStatistical inference was conducted using Random Field Theory . This usVisual observation of the neuromuscular activation patterns of the different quintiles showed differences in both muscle pair activation Figs  and 4 anThe neuromuscular activation patterns of the different quintiles showed that participants who landed with less peak knee flexion (red line) showed an increased activation of the {VM,HM} vector and the {HM,HL} vector during initial contact and the preparatory phase  mainly dWe also found significant associations between peak hip joint angles and neuromuscular activation patterns. Participants who landed with less peak hip flexion (red line) showed a higher activity of the {VM,VL} and the {VL,HL} vector during the peak loading-phase compared to other quintiles. A higher, more dominant activity of the VL was found in the {VM,VL} vector, this indicates a high activity of the VL in relation to the activity of the VM in the upper quintile during the peak loading phase . The incTables Canonical correlation analysis showed a significant relationship between the {peak knee flexion angle, peak hip flexion angle) vector and the {VM,VL,HM,HL} (time) vector field, we therefore further examined this relationship by considering combinations of EMG components with separate kinematic predictors. Additional CCA showed significant positive associations between peak knee joint flexion angle and the medial, lateral and posterior EMG vector of the quadriceps/hamstrings activation (p<0.05) indicating that muscular activation patterns of {VM,HM}, {VL,HL}, and {HM,HL} are significantly associated with a smaller (i.e. more erect) knee flexion angle during DVJ . More spAdditional 1-D SPM linear regression analyses revealed that the medial hamstring (HM) activity was significantly positively associated with the peak knee joint flexion angle suggesting that less peak knee flexion (i.e. a more extended knee) resulted in more HM activity during the preparatory and initial contact phase (p < 0.001) . In contAs can be seen in Additional 1-D SPM analyses showed a significant positive association between the VL activity during the peak loading phase of a DVJ and the peak hip joint flexion angle suggesting that athletes who perform DVJ with less peak hip joint flexion (i.e. a more extended hip) show a significantly higher VL activity during the peak loading phase  or vice The purpose of this study was to assess if sagittal plane landing kinematics of the knee and hip joints may predict neuromuscular activation patterns of quadriceps and hamstrings during the performance of DVJ. First, all subjects were divided into 5 quintiles based on their peak knee and hip flexion angles. The differences in neuromuscular activation, landing kinematics and kinetics were compared between the upper (flexed landing pattern) and lower quintile (erect landing pattern). Subsequently, correlation analyses were used to further assess the relationship between neuromuscular activation patterns and peak hip/knee joint flexion angles.As we made no hypothesis regarding a specific time point or muscle (pair) a priori, we analyzed the entire landing pattern  of four anatomically relevant muscle pairs . Subsequently, additional post-hoc 1-D SPM linear regression analyses between the individual muscles and knee/hip joint flexion angles were performed to help interpret the contribution of each individual muscle to the relation between muscle pairs and landing kinematics.The division of our sample into 5 quintiles based on peak knee and hip flexion angles Figs , 5 and 6The results of the correlation analyses further clarify the findings of the quintile analyses. A greater lateral EMG vector  during the peak loading phase (50–60% time), and greater medial and posterior EMG vectors  during the preparatory and initial contact phase (10–30% time) were found in participants who landed with less flexed knee joints . SimilarInterestingly, in contrast to recent literature  a decreaDespite the fact that we found an association between the knee flexion angle and the {VL,HL} vector during the peak loading phase, no relations were found between the individual muscle activations of neither the VL nor the HL and the knee joint kinematics . This su2 = 0.792) in peak external knee abduction moment in women. Decreased activation of VM and HM results in a diminished ability to resist external abduction loads. Another study [Interestingly, there was a time-shift when comparing the associations between peak knee flexion angles and the HM activity versus the associations between peak knee flexion angles and the {VL,HL} activation  . On the er study  showed tOn the other hand, participants who landed with less knee and hip flexion showed a significantly higher {VL,HL} vector during the peak loading phase. During this particular time phase of the landing, the {VM,VL} vector and VL amplitude were increased as well in the quintile that landed with less knee flexion. Previous studies ,17 demonInterestingly where we did find relations with the VL activity, no significant relations were found between the kinematic parameters and the individual VM activity. Recent work of Beaulieu et al.  showed tPrevious intervention studies have shown that movement re-education programs can successfully increase knee and hip joint flexion angles  and decrTo our knowledge, this study is the first to comprehensively reveal how neuromuscular activation patterns relate to the preferred landing pattern during the performance of DVJ. However the study still has some limitations that need to be taken into account. Firstly, as the human body acts as a linked-segment model, it might be important to implement more muscles and other joints than the knee and hip joint into the analyses. Less optimal movement patterns of the entire kinetic chain -including ankle, knee, hip and trunk- may contribute to ACL injury risk. Previous studies showed an influence of other proximal and distal musculature on the knee joint . FurtherThe current study has demonstrated clear associations during specific time periods between neuromuscular activation patterns and landing kinematics in the sagittal plane. The present findings have indicated that an erect landing pattern, characterized by less hip and knee flexion, was significantly associated with an increased activation of medial {VM, HM} and posterior {HM, HL} muscle pairs during the preparatory and initial contact phase and a more dominant and increased activation of anterior {VM, VL} and lateral {VL, HL} muscle pairs during the peak loading phase, respectively. Post-hoc analysis showed that an increased HM activation was mainly responsible for the increased medial and posterior activation during the preparatory and initial contact phase, and an increased VL activation was responsible for the increased anterior and lateral activation during peak loading. This suggests that participants landing in an erect pattern perform dynamic tasks with different neuromuscular activation patterns of the quadriceps/hamstrings complex. Future prospective studies should investigate if specific neuromuscular landing patterns are related to a higher ACL injury risk.S1 Appendix(PDF)Click here for additional data file.


The determinants of these species-associations and the adaptive significance of genetic divergence between host-associated viruses are poorly understood. One explanation is that epidemiological separation between reservoirs causes neutral genetic differentiation. Indeed, recent studies attributed host shifts to ecological factors and selection of ‘preadapted’ viral variants from the existing viral community. However, phenotypic differences between isolates and broad scale comparative and molecular evolutionary analyses indicate multiple barriers that Rabies virus must overcome through adaptation. This review assesses various lines of evidence and proposes a synthetic hypothesis for the respective roles of ecology and evolution in Rabies virus host shifts.Despite its ability to infect all mammals,  Current Opinion in Virology 2014, 8:68–72Virus evolutionThis review comes from a themed issue on Michael Brockhurst and Scott HensleyEdited by Issue and the EditorialFor a complete overview see the Available online 26th July 2014http://dx.doi.org/10.1016/j.coviro.2014.07.004http://creativecommons.org/licenses/by/3.0/).1879-6257/© 2014 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license (Rabies virus (RV) is a notorious multi-host pathogen that is capable of infecting all mammals, but paradoxically is maintained in distinct host species-associated transmission cycles, typically within the Carnivora and Chiroptera [iroptera . Howeveriroptera , 3. The The host-species association of RV is thought to arise from rare historical jumps into new species, which are followed by predominately within-species transmission . Such crin vivo and in vitro experiments, and recent rabies outbreaks resulting from cross-species transmission events, we assess whether ecological opportunity alone is necessary and sufficient for RV host shifts. We argue that broad scale historical patterns of cross-species transmission and host shifts, molecular evolutionary analyses of successful host shifts and phenotypic differences among host-associated RV variants all suggest a role for adaptive evolution in the establishment of new rabies reservoirs. We further propose a synthetic hypothesis for the respective roles of ecology and evolution in RV host shifts.The simplest hypothesis is that RV host shifts are determined purely by the ecological factors that provide opportunities for cross-species transmission and that genetic differences between host-associated variants arise from neutral evolutionary processes  variation in the degree of host-association among viral lineages due to variation in inter-specific contact rates among different reservoir hosts and (ii) that spill-over transmission and host shifts should be correlated with ecological overlap rather than the phylogenetic relatedness of host species. These predictions have been testable using large-scale phylogenetic analyses across many host species. Although certain reservoirs seem predisposed to infecting other species, true multi-host RVs (i.e. those that are maintained by multiple host species ) remain At the molecular level, evidence for positive selection in functional regions following host shift events would be suggestive of a role for adaptive evolution in RV host shifts. The nucleoprotein-encoding and glycoprotein-encoding genes  of RV isolates from various hosts are typically subject to strong purifying selection, with evidence for positive selection limited to a few G sites , 8, 19. Controlled experiments in which RVs are inoculated into atypical host species or cell lines offer further insights into the role of adaptive evolution in host shifts. Such studies have repeatedly demonstrated phenotypic differences among viral variants that may reflect optimisations to host ecological factors such as contact-rates to ensure sustained transmission (stage III and IV, If host adaptation is indeed the explanation for phenotypic differences among RV variants, one may expect a decreased ability to infect other species or patterns of clinical disease in incidental hosts that are less likely to lead to onward transmission. Although limited by low sample sizes, several heterologous host infection studies appear to demonstrate just that. When raccoons were inoculated with a raccoon-associated isolate or a dog-associated isolate, the homologous strain caused a long incubation period leading to acute clinical signs, whereas the heterologous dog strain caused only subtle neurological signs such as lethargy . These det al. [The effects of viral genotype on predisposition to host shifts are further illustrated through naturally occurring cross-species emergence events where detailed epidemiological surveillance has been coupled with viral sequencing. Across a series of outbreaks in striped skunks and gray foxes, Kuzmin et al. • found net al. •. Such ‘et al. •, suggeset al. [Another apparent example of pre-adaptation comes from a suspected host shift of skunk-associated RV into gray foxes. Borucki et al. •• showedet al. ••. This et al. ••.Taken together, all lines of evidence indicate that ecology alone is insufficient to explain the patterns of host shifts observed in RV. Although pre-adaptation may make some RV variants more likely to cross the species barrier than others, phenotypic differences and evidence of selection within recipient species suggest that this is not always sufficient. Once adaptation has occurred, it is maintained by ecological patterns of host-species associations, which provide ample opportunities for within-species transmission but far fewer opportunities for cross-species transmission. This ecological isolation sets the stage for purifying selection to maintain adaptive changes. This course of events may have great consequences on viral emergence. More reservoirs would enable exploration of more genomic space, increasing the likelihood of rare pre-adapted variants arising and potentially leading to a ‘snowball effect’ of ever increasing viral emergence. The importance of both pre-adaptation and post-emergence evolution may also explain the apparent paradox of constraints on host shifts, even to permissible hosts, despite the vast potential for rapid genetic change in RNA viruses. It could be that many potential host shifts were doomed from the beginning because of an inappropriate starting virus .Many questions regarding the adaptation of RV to hosts remain unanswered. For example, there is still no clear picture of the determinants of effective RV reservoirs. In this regard, a meta-analysis of the traits associated with known reservoirs may shed some light. The taxonomic scale of the effect of host phylogeny on emergence and establishment, the ecological or physiological mechanisms that constitute this barrier and the viral genotypic and phenotypic changes that overcome it also remain unknown. Finally, the seeming emergence of new, non-traditional reservoir species such as coatis , kinkajoin vitro or in vivo experiments. Synthesising ecological, virological and genomic studies provides a promising way forward to anticipate the fate of future RV emergence events and identify prospective new reservoir species.If we hope to understand RV adaptation, there is a clear need for full-genome sequencing studies, and perhaps even deep sequencing, to allow assessment of the roles of standing diversity and selection within donor and recipient hosts. Questions also remain regarding the repeatability of RV host shifts ••, which• of special interest•• of outstanding interestPapers of particular interest, published within the period of review, have been highlighted as:


Neural reflexes are stereotypical automatic responses often modulated by both intrinsic and environmental factors. We report herein that zebrafish larval C-shaped turning is modulated by the stimulated position of Rohon-Beard (RB) neurons. Targeted stimulation of more anterior RB neurons produces larger trunk flexion, which anticipates adult escape behavior by coordinated turning toward the appropriate direction. We also demonstrated that turning laterality varies with the numbers of stimulated neurons. Multi-cell stimulation of RB neurons elicits contralateral turning, as seen in the touch response to physical contact, while minimum input from single-cell stimulation induces ipsilateral turning, a phenomenon not previously reported. This ipsilateral response, but not the contralateral one, is impaired by transecting the ascending neural tract known as the dorsolateral fascicule (DLF), indicating that two, distinct neural circuits trigger these two responses. Our results suggest that RB neurons transmit the position and quantity of sensory information, which are then processed separately to modulate behavioral strength and to select turning laterality. Sensory-motor coordination adapts animal motilities to their perceiving environment. When aquatic vertebrates sense water vibration from an oncoming threat, they turn to the contralateral side by adopting a C-shaped body curvature of variable strength to elicit adequate escape trajectories. For example, sensory stimulus from the anterior evokes a large turning curvature to move toward the side opposite the threat, while stimulus from the posterior elicits small changes in direction to swim forward134Zebrafish larvae develop rapidly and acquire stereotyped motility patterns, which provide model networks for studying neural circuits developing from their conserved simple backbones. The first embryonic reflex is the touch response and is observed at 21 hpf (hours post-fertilization), whereby contact on the epidermis elicits body twists due to trunk muscle contraction7101012148In this study, we show that the turning response of zebrafish larvae is regulated according to their sensory receptive position. Specifically, RB neurons innervating a more-anterior region produce more extensive C-shaped body turning, suggesting the A-P position is perceived to control behavioral strength. We also demonstrate that stimulation of multiple RB neurons, rather than a single neuron, produce contralateral turning via intraspinal circuits. Nonetheless, stimulation of a single RB neuron elicits an ipsilateral turning response, which requires supraspinal circuits. Thus, our results suggest that sensory information from RB neurons is transmitted to supraspinal networks, while the robust escape response is triggered by intraspinal circuits that require multi-cell input from RB neurons.Transgenic zebrafish Tg(SAGFF36B/UAS:ChRWR-EGFP) express an optimized channelrhodpsin variant, ChRWR, in approximately 30% of RB neurons in a random and mosaic manner1719To our surprise, turning began from the ipsilateral side of stimulated neurons in 74 of the 90 typical responders, opposing the touch-response that begins with contralateral turning68Whereas the single-cell photostimulation elicited a unique feature in turning direction, the first turning angle varied depending on the A-P position of the neurons. When RB neurons were divided into three groups based on A-P level , stimuluThe results presented thus far indicate that turning direction varies in a stimulation mode-dependent manner such as photostimulation of single RB neurons eliciting ipsilateral turning, whereas stimulation of multicellular peripheral arbors or their epidermal contacts produces a contralateral response . We therFrom these results, we speculated that distinct sets of interneurons are activated in response to single- versus multi-cell stimulation. Studies of the zebrafish touch response suggested that reflex circuits are composed of spinal cord neurons, based on the turning response being maintained in lesion experiments following hindbrain removal. On the other hand, hindbrain RS neurons that play central roles in escape behavior of older fish are in place, and spinal cord neurons such as RB and CoPA extend their ascending axons to the hindbrain at the stage we examined the present study10118et al.81012815Studies of adult fish behavior generally described a tight correlation between the stimulus direction and initial body turning strength. Water vibration from the front evokes a larger turn than that from the back, as suited for avoidance trajectories in response to a threatIn this study, we showed that larval turning direction is switched in response to varied numbers of stimulated neurons. Multi-cell stimulation to RB neurons elicited turning to the contralateral side as seen in the touch-evoked response, and single-cell stimulation drove a novel type of turning ipsilateral to the stimulus. Epidermal sensory innervation is divided into trigeminal neurons on the head and RB neurons on the trunk, whereas the neural pathway from RB neurons is less understood. In the turning response evoked by the cranial sensory pathway, hindbrain Mauthner and other RS neurons are activated on the stimulated side, and their commissural descending projections excite spinal motor neurons directly and indirectly through descending interneurons152627288263029In contrast to the contralateral turning elicited by multi-cell stimulation of RB neurons, the ipsilateral response with single-cell stimulation exhibited different lesion susceptibility, indicating that a distinct circuitry involving supraspinal neurons executes the turning behavior. While the present study was limited in specifying all the neural components involved, the requirement of the opposite side of the DLF  implicatFish escape behavior is an immediate and robust startle reflex to significant threat, while harmless minor sensation or irrelevant noise should be filtered. In this regard, the requirement of multi-cell input for the robust intraspinal reflex makes sense in achieving escape by contralateral turning. What the ipsilateral response represents in larval zebrafish life remains an open question. For example, ipsilateral turning may help in capturing small planktonic animals that make minimum water vibration, or it may indicate cross talk with neural information for the A-P recognition, which might be suppressed in matured neural circuits. Although the current study is limited to the examination of early larval stages only, with transgenic ChRWR expression downregulated after those stages, it would be interesting to see if the ipsilateral response remains in later larval stages, juvenile, and adult zebrafish.Zebrafish RB neurons perceive mechanical sensation, and A-P positional information on the trunk region is processed for coordinating behavioral strength to take appropriate escape direction. Multi-cell stimulation on RB neuron is required for robust contralateral turning that is triggered by intraspinal reflex. On the other hand, less stimulation by single RB neuron elicits a novel type of ipsilateral turning, although the behavioral significance of this observation remains to be elucidated. Our results indicate that sensory input from RB neurons is processed differently for A-P recognition and triggering different types of behavior, through distinct neural circuits.Zebrafish were maintained in the laboratory fish room under a 14/10-h light/dark cycle. The larvae were maintained at 28.5 °C, with the developmental stages determined as previously described2, 1.67 mM HEPES, pH 7.2). Laser irradiation  or 333 fps . These sequential images were analyzed with Matlab  to measure maximum turning angle of the larvae between the midline and the tangent to the curved trunk surface at the proximal yolk tube level  agarose , and their trunks and tails were left to move freely in 1/3 diluted Ringer’s solution  and embedded in 2% LMP agarose. The neural tube was incised at the second-somite level using a razor blade under stereomicroscopy, and its complete disjunction was confirmed at the end of experiment following fixation in 4% paraformaldehyde . Transection of the DLF was performed under an upright microscopy  using a sharp glass capillary attached to a micromanipulator (Leica Microsystems GmbH). The DLF was labeled with GFP to guide the accurate incision and minimize artificial destruction of surrounding tissues.4 buffer  for 2 minutes, and then washed three times with PO4 buffer. GFP-positive RB cell bodies were punctured with a fine electrode and 10 mg/ml DiIC18(3)  in N,N-dimethylformamide  was iontophoretically injected. Larvae were fixed overnight at room temperature with 4% PFA, and then the extent of each labeled axon was measured using an all-in one fluorescence microscope .Anesthetized larvae were embedded in 1% agar, soaked in ice-cold 4% PFA in POHow to cite this article: Umeda, K. et al. Position- and quantity-dependent responses in zebrafish turning behavior. Sci. Rep.6, 27888; doi: 10.1038/srep27888 (2016).