During ET, mentally fatigued subjects reported higher RPE . ET induced a similar decrease in MVC torque , maximal voluntary activation level  and resting twitch  in both conditions. These findings reject our hypothesis and confirm previous findings that mental fatigue does not reduce the capacity of the central nervous system to recruit the working muscles. The negative effect of mental fatigue on perception of effort does not reflect a greater development of either central or peripheral fatigue. Consequently, mentally fatigued subjects are still able to perform maximal exercise, but they are experiencing an altered performance during submaximal exercise due to higher-than-normal perception of effort.It has been shown that the mental fatigue induced by prolonged self-regulation increases perception of effort and reduces performance during subsequent endurance exercise. However, the physiological mechanisms underlying these negative effects of mental fatigue are unclear. The primary aim of this study was to test the hypothesis that mental fatigue exacerbates central fatigue induced by whole-body endurance exercise. Twelve subjects performed 30 min of either an incongruent Stroop task to induce a condition of mental fatigue or a congruent Stroop task (control condition) in a random and counterbalanced order. Both cognitive tasks (CTs) were followed by a whole-body endurance task (ET) consisting of 6 min of cycling exercise at 80% of peak power output measured during a preliminary incremental test. Neuromuscular function of the knee extensors was assessed before and after CT, and after ET. Rating of perceived exertion (RPE) was measured during ET. Both CTs did not induce any decrease in maximal voluntary contraction (MVC) torque ( Self-regulation is the modulation of thought, affect, behavior, or attention via deliberate or automated use of cognitive control mechanisms  such as In the context of whole-body exercise physiology, we and others found that prolonged (30–90 min) engagement with CTs requiring self-regulation impairs endurance performance during subsequent running or cycling exercise . In thisAlthough the psychobiological model seems to provide a valid explanation for the negative effects of mental fatigue on endurance performance, at present we cannot totally exclude the possibility that the negative effects of mental fatigue on endurance performance may be mediated, at least in part, by the central component of muscle fatigue: central fatigue  post-CTs  compared to pre-CTs . However, neither the main effect of condition  nor the interaction  were significant. Vigor decreased  significantly after the self-regulation task (10.2 ± 3.0 to 8.3 ± 3.9) and the control task (10.6 ± 4.0 to 7.8 ± 4.7) with no significant difference between conditions .Self-reported fatigue was significantly higher  or time . Reaction time during both conditions did not change over time  but was significantly longer during the self-regulation task compared to the control task . Reaction time during the self-regulation task was significantly higher for all subjects.Response accuracy during CTs did not present any main effect of condition  but did not differ between conditions .Heart rate  = 0.031], time , or interaction .Cadence  during the whole-body ET did not present any main effect of condition . EMG RMS of the VL muscle was significantly higher during the mental fatigue condition compared to the control condition . EMG RMS of the RF muscle increased significantly during the whole-body ET  without any main effect of condition  or interaction . Time course of EMG RMS for the VL  and RF  muscles did not differ between conditions  and did not present a significant interaction .Electromyogram amplitude data are presented in Figure 4A) increased over time following both CTs . However, subjects rated a higher perceived exertion during the mental fatigue condition compared to the control condition . No significant interaction was demonstrated . Ratings of perceived exertion were significantly higher during the mental fatigue condition compared to the control condition for 9 out of all subjects .Rating of perceived exertion during the whole-body ET , none of NASA-TLX subscales presented any significant difference between conditions .Following the whole-body ET  = 0.032] nor interaction  on MVC torque of the knee extensors . Follow-up tests on the significant main effect of time  revealed that the CTs did not affect MVC torque . The whole-body ET caused a significant reduction in MVC torque in both conditions .There was no significant main effect of condition , doublet , Ct , and RFD  decreased significantly over time. The follow-up tests of the significant main effect of time are presented Table 1. M-wave amplitude of VL  and RF  muscles were not significantly affected by the CTs and the whole-body ET. M-wave amplitude of VL and RF muscles did not differ between conditions  and did not show any interaction .Peripheral parameters of neuromuscular function are presented in Figure 5. There was no significant main effect of condition  or interaction  on VAL . Follow-up tests of the significant main effect of time  revealed an increase in VAL post-CTs . On the contrary, the whole-body ET significantly reduced VAL . RMS/M-wave ratio of the VL muscle  did not present any significant main effect of time  or condition . Follow-up tests of the interaction  revealed that the RMS/M-wave ratio of the VL muscle decreased only during the control condition following the whole-body ET . RMS/M-wave ratio of the RF muscle did not change overtime  and did not present any main effect of condition  or interaction .Central parameters of neuromuscular function are presented in The primary aim of this study was to test the hypothesis that mental fatigue exacerbates central fatigue induced by whole-body endurance exercise. The results of the present study do not support this hypothesis. Furthermore, mental fatigue did not exacerbate peripheral fatigue induced by whole-body exercise. Therefore, the higher-than-normal perception of effort experienced by mentally fatigued subjects is independent of any central or peripheral alteration of neuromuscular function.We used a self-regulation task (incongruent Stroop task) to induce mental fatigue. The higher HR experienced during the incongruent Stroop task confirms that self-regulation is cognitively demanding and requires higher effort mobilization compared to the control task . The morInterestingly, both self-regulation and control tasks induced an increase in self-reported fatigue and a decrease in vigor suggesting presence of mental fatigue following both CTs. As in a previous study  a higherTo check that mental fatigue did not alter neuromuscular function at the onset of the whole-body ET, we performed neuromuscular function tests before and after the CTs. According to previous studies, completion of short  or proloInterestingly, as previously observed , the absIt has been suggested that CTs requiring self-regulation may cause the depletion of CNS resources, leading to reduced capacity of the CNS to recruit the working muscles , 2012. AMuscle fatigue can be caused by peripheral and/or central alterations  signals from premotor/motor to sensory areas of the cortex during voluntary muscle contractions (corollary discharge model; Despite that we did not measure intrinsic changes in the brain induced by prolonged self-regulation leading to mental fatigue, it is possible to speculate on the mechanisms involved based on previous studies. The ACC is strongly activated during incongruent Stroop tasks  and is aThis study was the first to test the hypothesis that mental fatigue and central fatigue induced by whole-body exercise are causally related. Contrary to this hypothesis, our findings show that mental fatigue does not exacerbate central fatigue during subsequent whole-body exercise. However, we must acknowledge some limitations. Firstly, the whole-body ET had to be performed on a cycle ergometer, inducing a time delay between the end of exercise and the start of neuromuscular testing due to the need to transfer the participant from the cycle ergometer to the isokinetic dynamometer. Therefore, the extent of muscle fatigue is likely to be underestimated in both experimental conditions. Secondly, the whole-body ET consisted of 6 min of high-intensity cycling exercise at a fixed workload. Future studies should investigate the effects of mental fatigue on more prolonged low-to-moderate intensity whole-body endurance exercise including running where the extent of central fatigue may be greater . The effDespite these limitations, this study provides further evidences that mental fatigue does not reduce the capacity of the CNS to recruit the working muscles. Our results suggest that the negative effect of mental fatigue on perception of effort does not reflect a greater development of either central or peripheral fatigue. Consequently, mentally fatigued subjects are still able to perform maximal exercise, but they are experiencing an altered performance during submaximal exercise due to higher-than-normal perception of effort. Therefore, further studies should investigate the brain alterations underlying the negative effect of mental fatigue on perception of effort and endurance performance. A better understanding of these brain alterations could lead to development of novel targeted interventions to decrease perception of effort and improve endurance performance in athletes, and reduced exertional fatigue in patients .The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


To examine the diagnostic performance of shorter versions of Alcohol Use Disorder Identification Test (AUDIT), including Alcohol Consumption (AUDIT-C), in identifying risky drinkers in primary care settings using conventional performance measures, supplemented by decision curve analysis and reclassification table.A cross-sectional study of adult males in general outpatient clinics in Hong Kong. The study included only patients who reported at least sometimes drinking alcoholic beverages. Timeline follow back alcohol consumption assessment method was used as the reference standard. A Chinese translated and validated 10-item AUDIT (Ch-AUDIT) was used as a screening tool of risky drinking.Of the participants, 21.7% were classified as risky drinkers. AUDIT-C has the best overall performance among the shorter versions of Ch-AUDIT. The AUC of AUDIT-C was comparable to Ch-AUDIT . Decision curve analysis revealed that when the threshold probability ranged from 15–30%, the AUDIT-C had a higher net-benefit than all other screens. AUDIT-C improved the reclassification of risky drinking when compared to Ch-AUDIT (net reclassification improvement = 0.167). The optimal cut-off of AUDIT-C was at ≥5.Given the rising levels of alcohol consumption in the Chinese regions, this Chinese translated 3-item instrument provides convenient and time-efficient risky drinking screening and may become an increasingly useful tool. The World Health Organization (WHO) indicated that alcohol consumption causes approximately 3.3 million net deaths globally in 2012, representing 5.9% of all global mortality , 5. A reThe limited time for consultations in primary care is a major constraint for more widespread use of risky drinking screening. Primary care physicians were more likely to use shorter screening tool for alcohol screening . In the The performance of abbreviated AUDIT as compared to its full version is traditionally determined on the basis of metrics such as sensitivity, specificity or area under the Curve (AUC). Despite the popularity of using these methods in diagnostic testing, there are increasing critics about their risk classification ability and their lack of clinical relevant –23. MostReclassification has recently become popular approach for comparing improvement among risk prediction models for diagnosing common diseases –28. A moa priori hypothesis that the shorter versions of Chinese language AUDIT had similar performance measures when compared with the full AUDIT.In China and Hong Kong, there has been interest in adopting greater screening and monitoring of drinking in clinical practice due to the rising rates of alcohol use , 4. In tTwo bilingual investigators preliminarily translated the original English instrument to the Cantonese version which was then examined for content validity by 15 native Cantonese speakers. The understanding and appropriateness of the wordings were discussed among the panel members. A modified Cantonese version was back-translated to English by another independent bilingual translator and this version was compared with the original English language AUDIT for equivalence. Based on the identified discrepancies, this process was repeated once more to produce a 10-item Chinese language that was subsequently pilot-tested on 28 male subjects who had reported ever consuming alcohol. Patients were then individually asked about the meaning of each item using a cognitive debriefing approach through face-to-face interviews. The investigators analyzed the pilot findings and a final 10-item Cantonese Chinese version was produced (Ch-AUDIT). Principal component analysis was used to investigate the psychometric properties of this final 10-item Ch-AUDIT. Similar to previous studies of AUDIT’s factor structure, the scree test indicated 3 factors could be extracted using criteria of Eigenvalue >1.0 and the total variance explained by these three components was 81%. The Ch-AUDIT also indicated good internal reliability .Data were collected by administering face-to-face interviews with a consecutive sample of 475 male patients attending public general outpatient clinics in Hong Kong. The general outpatient clinics are publicly funded primary care services , therefoAfter the completion of Ch-AUDIT interview, the participant’s alcohol consumption was assessed by the calendar-based timeline follow back (TLFB) approach. Using the TLFB approach, subjects will estimate the quantity of standard drink alcohol consumed on each of the 30 days preceding the interview. This approach serves as a validated reference standard as subjects estimated the amount of alcohol consumed on each of the 30 days preceding the interview . AverageR2 were used to assess the overall model performance. We used Hosmer-Lemeshow test to assess calibration of the prediction models by dividing subjects into deciles of risk and compared the predicted risk with the actual proportion of outcomes in each decile . Whereas, in this study the optimal cut-off point of AUDIT-C is ≥ 5, which might be explained by lower alcohol consumption in Hong Kong [Our findings are consistent with a meta-analysis of five cross-sectional studies comparing AUDIT-C against risky drinking in primary care setting (n = 8679) . The metong Kong . The lowong Kong . HoweverA cost effectiveness modeling study on UK health system highlighted that screening all primary care attendees with full AUDIT rather than AUDIT-C offers a slightly improved sensitivity, but the incremental cost-effectiveness ratio of £62,000 per QALY gained suggests that using the full questionnaire is not cost-effective . AnotherCompared to previous studies, a major strength of the present study was the utilization of more up-to-date performance measures to conduct head-to-head comparisons between the different AUDIT versions. However, this study has several limitations. First, we could only study one single aspect of problematic drinking, which is risky drinking behavior as defined by TLFB, and we could not conduct analysis on other alcohol problems, such as alcohol abuse or alcohol dependence. Nevertheless, the prevalence of alcohol abuse or dependence has traditionally been low in Hong Kong, as well as in China  . Second,This is the first study that compares the diagnostic performance between various shorter versions of AUDIT using decision curve analysis and reclassification table. The study sample included only male primary care patients who reported at least sometimes drinking alcoholic beverages. In conclusion, our findings confirm the validity of several short versions of the AUDIT, specifically AUDIT-C. This 3-item instrument provides a convenient and time-efficient screening of risky drinking in primary health care setting. We recommend a cut-off (≥5) score of AUDIT-C for populations with low prevalence of alcohol abuse and dependent, such as Hong Kong, to identify risky drinkers. Given the rising levels of alcohol consumption in the Chinese regions, the Chinese translated and validated abbreviated versions may become increasingly useful tools in primary care practice for reducing the harms of alcohol use.


Renal cell carcinoma with tumor thrombus extension into the inferior vena cava occurs in approximately 5% of cases. Despite such situations, an aggressive surgical approach is recommended. However, intraoperative prevention of pulmonary embolism by a fragmended tumor thrombus is necessary. To prevent pulmonary embolism, placement of a temporary suprarenal filter has been attempted, however, the precise placement of a temporary filter between the level of the hepatic vein and right atrium is not always easy because of its migration, tilting, and strut fracture. Here we report a method for early occlusion control of the intrapericardial inferior vena cava to prevent pulmonary embolism during nephrectomy in level II or III renal cell carcinoma tumor thrombus.Our first case was a 37-year-old Japanese man with left renal cell carcinoma extending into the inferior vena cava below the main hepatic vein (level II) and our second was a 75-year-old Japanese man with right renal cell carcinoma extending into the retrohepatic inferior vena cava at the main hepatic vein (level III). En block resection of the kidney and the tumor thrombus was performed with the aid of partial extracorporeal circulation; the postoperative course of both patients was uneventful.Control of intrapericardial inferior vena cava is a feasible method to prevent pulmonary embolism. Renal cell carcinoma with tumor thrombus extension into the inferior vena cava (IVC) occurs in approximately 5% of cases . DespiteThe patient was a 37-year-old Japanese man with a 10-cm left renal cell carcinoma extending into the retrohepatic IVC below the main hepatic vein (level II), as described by Naves and Zincke . Since aThe patient was a 75-year-old Japanese man with a 9.5-cm right renal cell carcinoma extending into the retrohepatic IVC at the main hepatic vein  .Written informed consent was obtained from both patients for publication of this Case Report and any accompanying images. A copy of the written consent is available for review by the Editor-in-Chief of this journal.


In this situation, statistical machine-learning methods, a.k.a. multivariate pattern analysis (MVPA), have received growing attention for their ability to generate stimulus response models in a data-driven fashion. However, machine-learning methods typically require large amounts of training data as well as computational resources. In the past, this has largely limited their application to fMRI experiments involving small sets of stimulus categories and small regions of interest in the brain. By contrast, the present study compares several classification algorithms known as Nearest Neighbor (NN), Gaussian Naïve Bayes (GNB), and (regularized) Linear Discriminant Analysis (LDA) in terms of their classification accuracy in discriminating the global fMRI response patterns evoked by a large number of naturalistic visual stimuli presented as a movie. Results show that LDA regularized by principal component analysis (PCA) achieved high classification accuracies, above 90% on average for single fMRI volumes acquired 2 s apart during a 300 s movie (chance level 0.7% = 2 s/300 s). The largest source of classification errors were autocorrelations in the BOLD signal compounded by the similarity of consecutive stimuli. All classifiers performed best when given input features from a large region of interest comprising around 25% of the voxels that responded significantly to the visual stimulus. Consistent with this, the most informative principal components represented widespread distributions of co-activated brain regions that were similar between subjects and may represent functional networks. In light of these results, the combination of naturalistic movie stimuli and classification analysis in fMRI experiments may prove to be a sensitive tool for the assessment of changes in natural cognitive processes under experimental manipulation.Naturalistic stimuli like movies evoke complex perceptual processes, which are of great interest in the study of human cognition by functional MRI (fMRI). However, conventional fMRI analysis based on  In spite of this success, the statistical analysis and interpretation of BOLD fMRI data with the goal of understanding brain function remains a daunting task that continuously motivates the development of new and improved analytical methods. Conventional hypothesis-driven fMRI analysis of task-evoked activity involves an explicit model of the BOLD fMRI response to the stimulus. Relevant stimulus features must be identified a priori and transformed by an equally predetermined hemodynamic response function (HRF) to form the design matrix of a general linear model (GLM). By contrast, data-driven machine-learning (ML) methods, often referred to as multivariate pattern analysis (MVPA) in the context of fMRI, are attractive, because they do not require any generative model of the BOLD signal. ML algorithms for clustering, decoding, and especially for classification facilitate the quantification of stimulus-related information without making strong assumptions about the nature of the BOLD fMRI signal . This is not implausible, since attention is known to have a strong modulatory effect in cognitive experiments , linear discriminant analysis (LDA), and Gaussian Naïve Bayes (GNB)] for the analysis of fMRI data, but all of those focused on binary classification, a small number of stimulus categories such as houses and faces, and input from small predefined brain regions of just a few hundred voxels .In the recent literature, fMRI data from movie-viewing experiments have mostly been analyzed by conventional (univariate) GLM analysis, which quantifies correlations of the fMRI signal with scalar regressors derived from visual features such as optical flow (motion), luminance or the presence of faces in the movie  and a large number of more or less distinctive classes (>100 stimulus time points) that far exceeds the number of experimental repetitions (< 10 stimulus presentations). In this work, several suitable classification methods were therefore systematically evaluated to find a maximum in classification accuracy, which is a proxy for the information content of the fMRI signal. Given the conceptual proximity between classification methods and clustering algorithms this line of research may eventually bridge the gap between data-driven and hypothesis-driven fMRI analysis by revealing the relevant stimulus features underlying the fMRI signal in natural conditions.Four subjects  each gave written informed consent in accordance with institutional guidelines at the NIH (IRB-approved protocol 00-N-0082) to participate in a series of fMRI experiments. During each fMRI experiment the iconic opening scene from the popular action movie “The Matrix” was presented for exactly 5 min (video without audio). This kind of naturalistic stimulus was intended to elicit a broad range of perceptual and cognitive processes thus precipitating a varied fMRI response, suitable for analysis by machine-learning (ML) classification algorithms. Experiments were repeated over 4–6 separate scan sessions spread over several months until each subject had completed eight viewings of the same video stimuli, 1–2 per session depending on time constraints.MRI experiments were performed on a high-field 7 T MRI scanner  equipped with 70 mT/m @ 200 T/m/s gradients and a combined 2-channel transmit 32-channel receive head coil . http://psychtoolbox.org) in Matlab served to play the digital video stimuli in synchrony with MRI acquisition triggers and the eye tracker. In addition, each subject's cardiac and respiratory signals were recorded by pulse-oxymetry and a pneumatic chest belt.A 3 cm gap between the outer transmit and inner receive coil accommodated a mirror and a small MR-compatible CCD camera  to allow both the projection of visual stimuli and video recording of the subject's eye movements using eye-tracking software by Arrington Research, Inc. (USA). A separate computer running the Psychophysics Toolbox  as a separate stimulus category for classification, even though some stimuli might be more similar than others in terms of the evoked BOLD fMRI response. Unlike other studies, this work was not aimed at determining what visual features drive the fMRI response and how the brain might categorize stimuli. Instead classification accuracy, i.e., the percentage of correctly identified test stimuli, served us purely as a measure of information extracted from the fMRI signal and therefore classifier performance.Previous studies of fMRI during movie stimulation have mostly focused on either inter-trial correlations as a measure of reproducibility  that are most informative for classification, we repeated all classification analyses for feature sets ranging from 16 to 216 voxels, selecting those that were most significantly modulated by the stimulus. The relative significance (or fMRI activation strength) in each voxel location was estimated by a univariate analysis of variance (ANOVA), treating each of the 150 fMRI volume acquisitions during the movie as an independent group (or factor) and each experimental run as a repeated measurement  or nearest-mean (NM) classification scheme and evaluated by leave-one-experiment-out cross-validation. In other words, any sample pattern (from the test data) is assigned the same class as its nearest neighbor from a set of class templates (from the training data) as determined by a metric of pairwise distances. Because systematic differences between fMRI scans constitute a large part of the noise confounding classification, the cross-validation scheme strictly avoided sharing data from any one experiment between training and validation data. Without this leave-one-experiment-out cross-validation scheme, scan specific variations e.g., in shimming, coil sensitivities, subject performance or even breathing patterns are likely to outweigh inter-stimulus differences especially for similar stimuli close in time. In other words, different samples (classes) from the same fMRI scan were often more similar than samples from the same class in different scans.The multi-class classifiers compared here are in essence all based on the efficient Table centroid) and a covariance matrix estimated from the residuals of its members (after subtracting the class mean). Variants of discriminant analysis are typically qualified as linear or quadratic as well as GNB, all of which refer to constraints imposed on the estimated model covariance matrix: Linear and quadratic discriminant analysis respectively refer to a common (pooled) or a class-specific (stratified) estimate of the within-class (noise) covariance matrix. Similarly, the GNB classifier assumes a diagonal noise covariance matrix —a simplifying assumption that is not necessarily accurate, but convenient and often surprisingly effective in situations where a small number of samples in a high-dimensional input space renders the covariance matrix rank deficient and not invertible.The simplest classifier tested here can be described as pairwise nearest-neighbor classification with each (single) experimental run in turn serving as a set of class templates (training data), against which every fMRI volume from any other experimental run (test data) was compared either by the Euclidean or the correlation distance , which was employed here as a form of regularization to facilitate LDA based on the Mahalanobis distance that requires a full-rank covariance matrix. All the nearest-mean LDA-type classifiers were tested by leave-one-experiment-out cross-validation, meaning that PCA, class centroids and covariance matrices were computed from training data that included all but one experimental run, while the left-out experiment served as validation data for testing classification results. The comparisons in Figures after averaging over experiments. An alternative computation of PCA on training data concatenated in time resulted in a much larger number of eigenvectors, some apparently specific to individual experiments, others obviously non-neuronal . To compute sample distances and classification results, both the training and validation data were projected into the spatial PCA eigenbasis of the training data. Note that the resulting signal time series are not normalized in variance, unlike the temporal PCA eigenbasis. Figure 15 voxels in each of the four subjects. For display purposes only, the temporal PCs estimated from the fROI were projected back to the whole fMRI volume, such that any correlations outside the fROI would also be observed.Another common solution to this The eye-tracking data confirm a large degree of consistency in the way subjects watch the movie stimuli, notwithstanding a considerable amount of variance due to technical limitations. Eye blinks, head motion and subject non-compliance, i.e., fluctuations in vigilance, are major sources of variability between experimental runs and subjects. Obvious manifestations of subject drowsiness are extended intervals of partial or complete eye closure and corresponding periods of fMRI signal that is inconsistent with other experimental runs. As a result, subjects, who were less able to maintain vigilance over a number of experimental runs, showed lower average classification accuracy. Nevertheless, after removing obvious artifacts  the eye traces recorded at 30 Hz exhibit a high degree of correlation across experimental runs and between subjects.  In fact, an overlay of gaze positions from multiple experiments onto the dynamic movie stimulus makes it obvious that there are rarely more than one or two foci of attention at any given moment during the movie.ventral visual pathway of high-level object recognition. Functional ROIs of adjustable size were defined on these maps by selecting between 16 and 216  of the most activated voxels, i.e., those showing the highest F-values. For reasons of computational efficiency, this input ROI for classification analysis was confined to the occipital half of the fMRI volume, where virtually all the stimulus-correlated fMRI signal was concentrated. By contrast, frontal regions close to the paranasal sinuses were strongly affected by susceptibility artifacts as well as a stimulus-correlated motion artifact in the eyes, which corroborates the consistent eye-tracking results and would certainly confound stimulus classification.The ANOVA F-statistic maps from a typical subject Figure  clearly The main results of this study are summarized in Figure A similar dome-shaped dependence was observed when testing classification rates as a function of dimensionality reduction using 8–150 principal components . By comparison, the nearest-mean classifiers , the LDA and GNB classifiers based on Dimensionality reduction by PCA in combination with GNB or LDA strongly increased classification rates in our experiment. It was therefore of interest to investigate the spatial basis functions that account for most of the signal variance as well as their within-class noise covariance. Both of these determine each component's weighting as part of the normalized Euclidean and the Mahalanobis distance metrics. The basis functions computed by PCA can be viewed as spatial brain maps Figure  that indbetween-class over within-class variance for each temporal PC , it is reassuring to note that all of the leading PCs reflect the known functional anatomy of the visual cortex and other functionally related brain regions. Notably, there is a first PC common to all subjects that broadly covers lateral occipital and ventral temporal cortices. The second common component highlights the motion sensitive area V5/hMT and has the highest F-statistic in all subjects. Many of the following PCs  seem to contrast different sub-regions of the visual cortex V1-4, which one would expect to be represented by a number of PCs. In general, the leading PCs exhibit a clustered structure of low spatial frequency and bilateral symmetry that resembles functional networks reported in studies of task-based and resting-state functional connectivity  for the purpose of reliably eliciting a widespread, diverse, and reproducible BOLD fMRI signal response in human subjects. Consistent with previous studies, the naturalistic movie stimuli evoked reproducible activity in the occipital and ventral temporal cortex  before declining again with higher voxel counts. The rise, peak and fall of classification rates as a concave function of functional ROI size is more than plausible under the assumption that the voxel features added first are highly informative, while those added later are apt to confound classification with a higher noise contribution and less signal reproducibility as evidenced by lower values of the F-statistic. Since all classifiers consistently benefited from a large fROI one may also conclude that stimulus-related information in the brain is spatially distributed and its coarse global structure most reliably interpreted.Previous fMRI studies have applied and compared classification methods mostly based on experiments with few distinct stimulus categories presented in a block-design or event-related paradigm  as templates  confirmed that classification accuracy varied and tended to be higher when training and validation data were acquired during the same scan session Figure : In spitintra-class noise variance. This variance normalization made the number of PCs another critical optimization parameter, as results deteriorated again with an increasing number of low-variance PCs  signal and the residuals (after subtraction of the class means). Note that PCA alone did not confer any increase in classification accuracy, although it did offer a 100-fold reduction in dimensionality ~64/16,000) without substantially compromising results  feature-selection algorithms is beyond the scope of this paper that focuses on a comparison of classification methods.between subjects; such hyperalignment methods are not directly pertinent to this study  signal sources Figure . One migThe first PC in all subjects reflects a global unison activation of cortical regions accounting for the largest fraction of variance but not the highest F-statistic. This PC may well reflect a system-wide modulatory effect like arousal. The second PC exhibits the highest F-statistic indicating the most reproducible stimulus response in all subjects. This and the apparent involvement of motion-sensitive areas V5/hMT suggests that PC#2 reflects optical flow and motion, which are known to account for a large proportion of fMRI signal variance throughout the visual system  generated by a preceding GLM analysis, which is sometimes termed an encoding model and constitutes a form of averaging that constrains the ML model .The present study confirms the suitability of several classification methods for fMRI experiments with movie stimuli and it demonstrates the separate influences of the distance metric and the feature space (fROI) on absolute classification rates. In analogy to previous work  perceptual processes by fMRI. They evoke a diverse yet robust and reproducible fMRI response throughout large visual areas in the lower occipital and temporal cortices. In a comparison of common classifiers average classification accuracies above 90% were achieved by the most successful combination of trial averaging, PCA and LDA. This is remarkably high, considering the relatively large number of classes (150) used in this study. However, absolute classification rates are not too meaningful, since they depend on many factors such as feature selection, variance normalization and other pre-processing of the data. Nevertheless, high classification rates may translate into high sensitivity for the detection of various cognitive and perceptual processes. The optimization of classification algorithms also reveals properties of the fMRI signal: Best classification results are supported by a global fROI, indicating that information about visual stimulus identity is not local but distributed throughout occipital and temporal cortices. Interestingly, the spatial and temporal patterns of the dominant PCA components are similar between subjects and reminiscent of functional connectivity networks.The results presented above demonstrate how fMRI experiments with repeated naturalistic movie stimuli can suitably be analyzed using multivariate classification methods tailored to data with high dimensionality, numerous categories (and few repetitions). Cinematographic motion pictures are convenient HM, Design of study and experimental procedures, data acquisition and analysis, interpretation of results, preparation of manuscript and figures. JDZ, Experimental design and data acquisition, interpretation of results, preparation of manuscript. JD, Study design, interpretation of results, and manuscript preparation.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Withania somnifera Dunal, is one of the most commonly used medicinal plant in Ayurvedic and indigenous medicine traditionally owing to its therapeutic potential, because of major chemical constituents, withanolides. Withanolide biosynthesis requires the activities of several enzymes in vivo. Cycloartenol synthase (CAS) is an important enzyme in the withanolide biosynthetic pathway, catalyzing cyclization of 2, 3 oxidosqualene into cycloartenol. In the present study, we have cloned full-length WsCAS from Withania somnifera by homology-based PCR method. For gene function investigation, we constructed three RNAi gene-silencing constructs in backbone of RNAi vector pGSA and a full-length over-expression construct. These constructs were transformed in Agrobacterium strain GV3101 for plant transformation in W. somnifera. Molecular and metabolite analysis was performed in putative Withania transformants. The PCR and Southern blot results showed the genomic integration of these RNAi and overexpression construct(s) in Withania genome. The qRT-PCR analysis showed that the expression of WsCAS gene was considerably downregulated in stable transgenic silenced Withania lines compared with the non-transformed control and HPLC analysis showed that withanolide content was greatly reduced in silenced lines. Transgenic plants over expressing CAS gene displayed enhanced level of CAS transcript and withanolide content compared to non-transformed controls. This work is the first full proof report of functional validation of any metabolic pathway gene in W. somnifera at whole plant level as per our knowledge and it will be further useful to understand the regulatory role of different genes involved in the biosynthesis of withanolides. Withania somnifera (Indian ginseng) is one of the most famous and widely used medicinal plants  belongs to the 2–3 oxidosqualene cyclases (OCSs) gene family along with α-amyrin synthase (AAS), β-amyrin synthase (BAS), lupeol synthase (LS), and lanosterol synthase (LAS) –14. Cyclomnifera . CAS is thaliana  Glycyrrha glabra , Pisum s sativum , Rhizoph stylosa , Kandelia candel  are alsoa candel . Anotherataricus . Though omnifera –30. Theiomnifera –32. Tranomnifera –34.via transgenic approach two methods are very effective one of which involves loss of gene function i.e. RNAi mediated mechanism and other through gain of function or over-expression approaches. Both of these mechanisms together serve as an important tool to validate the function of a geneRNA-mediated gene silencing is initiated by the delivery of double-stranded RNA (dsRNA), which is processed into small interference RNA molecules (siRNA) of 21–25 bp by Dicer (DCR), which depends on ATP and RNaseIII. The siRNA is then incorporated into the RNAi-induced silencing complex (RISC), which in turn degrades the target mRNA . Our findings suggest that expression level of WsCAS was modulated in response to heat cold, wound, MeJA, and SA treatments in condition specific manner. HPLC analysis showed the relative change in withanolide content also  serve as key enzymes in secondary metabolite synthesis by driving the metabolic pool of isoprenoids and converting it to a range ofhigher terpenoid- secondary metabolites squalene –14. Cycl studied –20. It hormation . Secondaynthesis . A simillicitors . Modulatent also . These fin planta. Transgenic approaches provide a powerful tool for metabolic engineering and gene function studies in Withania plants. Based on the improved transformation system and development of frequent changes and progression in the field of RNAi, we carried out the study on the gene function investigation by RNAi technology. RNAi mediated gene silencing studies was an effective measure to carry out gene function investigation and further improvement in crop plant species [Camellia sinensis [Panax ginseng, significant reduction in transcript level and subsequently ginsenoside production was observed [WsCAS gene spanning various regions of the gene  of full length WsCAS  or XbaI (overexpressing lines) digested genomic DNA from the shoot of T0 transgenic plants, and probed with a DIG-labeled DNA probe of partial PCR amplified fragment of bar/nptII gene. The results showed the bar/nptII sequence was present in all T0 transgenic plants analyzed, and mostly each line contained single integrated loci of the bar/nptII gene  gave a higher silencing efficiency in range from 93.3 to 98.9% than the middle (WsRNAi 2) and 3’ IR (WsRNAi 3) construct. Withanolide content from the transformed plants was analyzed by HPLC analysis. Several silenced lines with low levels of withanolide content were obtained, in which, withanolide content decreased considerably showing the down regulation mediated by RNAi of WsCAS. The results demonstrated that the withanolide content of transformants differed significantly in silenced lines with varying levels of individual withanolide as compared to the untransformed one. Manipulation of saponin biosynthesis by RNA interference-mediated silencing of b -amyrin synthase gene in soybean exhibited a stable reduction in seed saponin content, correlated with the b -amyrin synthase mRNA depletion [WsCAS gene expression was due to the formation of the WsCAS RNA hairpin in vivo, which resulted in a decrease in WsCAS enzyme activity, ultimately resulting in a reduction in the biosynthesis of withanolides  and a structural gene (G10H) involved in MIA pathway increased the accumulation of monoterpenoid indole alkaloids [The results showed that the transcription level of epletion . In tomaepletion . Similarepletion . From thanolides . In the formants . In Cathlkaloids . Overexplkaloids .Withania. However variation in direct linkage between CAS transcript level and relative withanolide content was observed in transgenic lines. The reason for that withanolide is not the immediate product of cycloartenol synthase. The withanolide biosynthesis involves various post-transcriptional and post- translation modification steps [. Studies on biosynthetic pathways and metabolic engineering already showed that the fluxes through the pathways are controlled not only by gene expression levels, but also by post-translational regulation of enzyme activity and enzyme and metabolite compartmentalization and transport [Therefore, inverted repeat silencing construct under the control of the constitutive promoter were able to effectively down-regulate the pathway gene expression in on steps . Differeon steps . Studiesransport . Also thW. somnifera and also generating over-expressed lines with elevated levels of withanolides. Strategies utilized in both the experiments were different but it complemented with each other in the sense that both helped in determining the function of the WsCAS gene. The observed decrease in WsCAS transcription level in Withania silenced lines very likely resulted in a reduction in WsCAS enzyme activity in vivo, which in turn responsible for reduction in the biosynthesis of withanolides by regulationg a key position in the biosynthetic pathway by WsCAS.In conclusion, the present study was an attempt to generate transgenic lines containing a construct which impairs the normal functioning of a key pathway enzyme cycloartenol synthase (CAS) thereby suppressing withanolide biosynthesis in S1 Fig(TIF)Click here for additional data file.S2 Fig(TIF)Click here for additional data file.S3 Fig(a) silencing lines. (b) over expressing lines.(TIF)Click here for additional data file.S1 Table(DOC)Click here for additional data file.


A complete database of the mechanical properties of an epoxy polymer reinforced with uni-weave carbon fibre non-crimp fabric (NCF) is established. In-plane and through-the-thickness tests were performed on unidirectional laminates under normal loading and shear loading. The response under cyclic shear loading was also measured. The material has been characterised in terms of stiffness, strength, and failure features for the different loading cases. The critical energy release rates associated with different failure modes in the material were measured from interlaminar and translaminar fracture toughness tests. The stress–strain data of the tensile, compressive, and shear test specimens are included. The load–deflection data for all fracture toughness tests are also included. The database can be used in the development and validation of analytical and numerical models of fibre reinforced plastics (FRPs), in particular FRPs with NCF reinforcements. Specifications Table•This data set presents a complete mechanical characterisation of a CFRP system.•The data can be used as input properties in analytical models.•The data can be used as input parameters in finite element analyses and used for validation of results.•The data can be compared to already available data for others CFRPs. The data can also be used in the development of future CFRPs, in particular those with NCF reinforcements.•Guidelines for the mechanical and fracture characterisation of a given FRP material are provided.1•in-plane longitudinal tension•in-plane longitudinal compression•in-plane transverse tension•in-plane transverse compression•through-the-thickness (TT) tension•TT compression•in-plane shear•TT shearThe stress–strain curves under the following loading cases are presented:The following terminology is used: 1-index refers to the longitudinal (to the fibre) direction in the reinforcement plane, 2-index refers to the transverse direction in the reinforcement plane, and 3-index refers to the TT direction w.r.t. the reinforcement plane. The stiffness and strength values are extracted from the stress–strain curves, and the specimen failure features reported.R-curves).Load–deflection curves are obtained from interlaminar fracture toughness tests in mode I, mode II and mixed-mode, and from translaminar fracture toughness tests. The energy release rates associated with the initiation of crack growth for the different tests are reported, as well as the crack resistance curves (The dimensions of the tests specimens are reported in The carbon fibre reinforced plastic (CFRP) material system is an HTS45/LY556. The Hunstman LY556 epoxy resin was supplied by ABIC Kemi AB. The reinforcement layer is a 205 GSM uni-weave non-crimp fabric (NCF), from Porcher Industries. It consists of HTS45 E23 Tenax® carbon fibre bundles, which are held together by glass fibre/polyamide weft threads . HTS45/L3.1The test procedure for the tensile and compressive in-plane tests followed the ASTM standard D 3039 tion Eq. . Only thLongitudinal tensile specimens exhibited broom-like fracture, 3.2Iosipescu tests, documented with the ASTM standard D 5379 The failure mode of the Iosipescu specimens was premature failure at the notches by splitting, followed by shear failure in the gauge section . This fa3.3Double cantilever beam (DCB), end notched flexure (ENF) and mixed-mode bending (MMB) interlaminar fracture toughness tests are documented by test method standards R-curves, in R-curves in 2.The critical energy release rates The fracture surfaces of DCB, ENF and MMB specimens were not perfectly flat but exhibited some waviness, which is specific of textile FRPs . The for3.4The TT tensile and compressive data were extracted using the double waisted specimen design proposed by Ferguson et al. For the compressive tests, the specimens were simply loaded between two parallel platens in displacement control equivalent to an initial strain rate of approximately 2%/min. Back-to-back strain measurements and stereo DIC measurements indicated no specimen bending. The strains were averaged from the DIC measurements over the entire surface of constant gauge section. The surface monitored by the DIC system was not always the same in all specimens so that the evaluation of both Poisson׳s ratios For the tensile loading configuration, rod end bearings were attached to the universal testing machine to prevent the introduction of moments in the specimens. The specimen end surfaces were adhesively bonded to two steel plates connected to the bearings. Strain gauges were bonded at the centre of the wider surfaces of the specimen, and the average of the two strain readings was considered to construct the stress–strain curves. In two specimens, the strain gauges produced inaccurate signals and the strain data were discarded. However, the strength values associated with these two specimens are considered reliable.3.5The test procedure described by Pinho et al. J-integral of the specimen configuration (geometry and layup considered) with finite element methods.Cross-ply specimens are needed to prevent splitting at the notch when the crack initiates. The data reduction scheme, based on Eqs. (1)From the critical energy release rate for the laminate, the critical energy release rate for the 0°-plies is found using Eqs.


The method requires arterial blood sampling from a radial artery , and the aim of this study was to develop a method for extracting an image-derived, non-invasive input function from a volume of interest (VOI).Metabolic liver function can be measured by dynamic PET/CT with the radio-labelled galactose-analogue 2-[18F-FDGal PET/CT data from 16 subjects without liver disease  and 16 patients with liver cirrhosis were included in the study. Five different input VOIs were tested: four in the abdominal aorta and one in the left ventricle of the heart. Arterial input function from manual blood sampling was available for all subjects. K*-values were calculated using time-activity curves (TACs) from each VOI as input and compared to the K-value calculated using arterial blood samples as input. Each input VOI was tested on PET data reconstructed with and without resolution modelling.Dynamic K*-values that correlated significantly with K calculated using arterial blood samples. Furthermore, TACs from two different VOIs yielded K*-values that did not statistically deviate from K calculated using arterial blood samples. A semicircle drawn in the posterior part of the abdominal aorta was the only VOI that was successful for both healthy subjects and patients as well as for PET data reconstructed with and without resolution modelling.All five image-derived input VOIs yielded 18F-FDGal PET/CT can be measured without arterial blood samples by using input data from a semicircle VOI drawn in the posterior part of the abdominal aorta.Metabolic liver function using  One method for defining an image-derived input function for 18F-FDG of the liver was published some years ago with a circular Aorta-VOI drawn in the middle of the abdominal aorta for 18F-FDG PET of liver tumours using a dedicated PET camera [Because T camera . It is r18F-FDGal PET/CT. This makes the method safer, simpler, and more generally applicable for routine clinical and scientific use.We have defined an image-derived input function in terms of a semicircle drawn in the posterior part of the abdominal aorta that could replace arterial blood sampling successfully for quantification of metabolic liver function using


Alveoli, the milk-producing units of the mammary gland, are generated during pregnancy by collaboration of different epithelial cell types. We present the first analysis of transcriptional changes within the hormone sensing population during pregnancy. Hormone-receptor positive (HR+) cells play a key role in the initiation of alveologenesis as they sense systemic hormonal changes and translate these into local instructions for neighboring HR- cells. We recently showed that IGF2 is produced specifically by HR+ cells in early pregnancy, but is undetectable in the virgin state. Here, we define the transcriptome of HR+ cells in early pregnancy with the aim to elucidate additional changes that are unique for this dynamic developmental time window.We harvested mammary glands from virgin, 3-day and 7-day pregnant mice and isolated a few hundred hormone-sensing cells per animal by FACS for microarray analysis. There was a high concordance between animals with a clear induction of cell cycle progression genes at day 3 of pregnancy and molecules involved in paracrine signalling at day 7.These findings underscore the proliferative capacity of HR+ cells upon specific stimuli and elucidate developmentally-restricted changes in cellular communication. Since the majority of breast cancers are HR+, with a variable proportion of HR+ cells per tumor, we anticipate that this data set will aid further studies into the regulation of HR+ cell proliferation and the role of heterotypic signalling within tumors.The online version of this article (doi:10.1186/s12861-015-0058-9) contains supplementary material, which is available to authorized users. The Pi score was used to determine the top 25 up- and down-regulated genes in the 3-day and 7-day pregnant samples compared to the virgin group. Principal components analysis was conducted on the expression covariance matrix to identify potential sample outliers . The microarray data from this publication have been submitted to the NCBI Gene Expression Omnibus and are deposited as GSE63720.Microarray hybridization and scanning was performed by the Genomic and RNA Profiling Core Facility at DUKE-NUS Graduate Medical School. 1.5 μg of biotinylated a/cRNA from each sample was hybridized at 58°C for 16 hours using the Illumina Whole-Genome Gene Expression Direct Hybridization Assay system with the Illumina Mouse WG-6 v2.0 (six-sample BeadChip) platform. The signal was developed using streptavidin-Cy3 and the BeadChips were scanned with an Illumina BeadArray Reader. Raw signals were logarithmically transformed (to base 2) and quantile normalized. The background noise from the array was determined at 100, and probes with an average signal <100 in all comparator groups were removed from further analysis . Differential gene expression between pregnant and virgin samples was expressed as a log ratio of the average logged signals between the compared groups. Statistical significance of differential gene expression was assessed by a regularized Cyber-T, ). False Cyber-T, . GeneralCyber-T,  and compwww.ingenuity.com). A pre-filtered list of differentially expressed genes (absolute fold-change ≥ 2-fold and FDR ≤ 10%) were used as input for each comparison. Pathway enrichment was assessed on the list of canonical pathways from the Ingenuity Knowledge Base. Over-representation of biological pathways was ascertained via Fisher’s exact test and corrected for multiple testing by the Benjamini-Hochberg procedure.Pathway enrichment analysis was conducted via the over-representation analysis (ORA) method in the Ingenuity Pathway Analysis (IPA) tool (Ingenuity Systems, All procedures were performed in accordance with the guidelines for ethical treatment of laboratory animals approved by the SingHealth Institute Animal Care and Use Committee, Singapore. FVB/N mice were purchased from Jackson Laboratory and bred and maintained in the animal facility of the DUKE-NUS Graduate Medical School and The National Cancer Centre, Singapore under guidelines for housing and husbandry conditions of each institute. Mice were euthanized by carbon dioxide inhalation and cervical dislocation. Effort was taken to ensure good animal welfare and prevent suffering.The data set supporting the results from this article is available in the NCBI Gene Expression Omnibus, GSE63720.


R) genes identified against plant viruses and the corresponding avirulence (Avr) genes identified so far. The most common models to explain the mode of action of dominant R genes will be presented. Finally, in brief the hypersensitive response (HR) and extreme resistance (ER), and the functional and structural similarity of R genes to sensors of innate immunity in mammalian cell systems will be described.To establish a successful infection plant viruses have to overcome a defense system composed of several layers. This review will overview the various strategies plants employ to combat viral infections with main emphasis on the current status of single dominant resistance ( During the past decades it has become clear that plants have a unique and complex defense system that consists of several layers, which enables them to avoid, suppress, or actively defend against pathogens from all kingdoms like fungi, bacteria, nematodes, and viruses. Of all plant viruses known, only a few cause serious diseases and, if so, mostly limited to a very small number of crops. In general, most viruses have a limited  host range and the number of so-called non-hosts exceeds those of hosts. In those plants that are hosts, viruses encounter different mechanisms of defense. Some act general against all viruses and this response is part of the innate immune system, while others are virus-specific and involve resistance genes. Triggering of the latter simultaneously mediates a rapid necrosis at the site of virus entry and prevents further spread of the virus throughout the host. In several cases resistance genes do not confer absolute resistance and low levels of virus replication can still be observed. In those cases the genes are referred to as partial resistance genes or tolerance genes.While throughout the years reviews on resistance genes have appeared with regular intervals, these mostly had their main focus on fungal and bacterial resistance genes, primarily due to the large amount of data available. This review aims to present an overview on the current status of resistance genes against plant viruses, with emphasis on single dominant resistance genes. The very basis of plant pathogens (among others plant viruses) not being able to infect all plants is due to a mechanism called non-host resistance (NHR)  and in the very early days post-transcriptional gene silencing (PTSG)]. RNA silencing is a host response triggered by double stranded (ds)RNA. These molecules thus act as a MAMP/PAMP and in which RNAi can be regarded as PTI. The main difference with pathogens such as fungi and bacteria is that recognition of viral MAMPs/PAMPs occurs intracellularly RNAs, and one of the hallmarks for antiviral RNAi, and the second one is that of (host-gene encoded) micro (mi)RNAs involved in gene regulation. The antiviral RNAi response is induced by viral double stranded (ds)RNA molecules that arise from replicative intermediates or secondary RNA folding structures. These structures are sensed by a host RNase type III-like enzyme called Dicer-like (DCL) protein and cleaved into short interfering (si)RNA of 21–24 nucleotides (nt) in size  This gene encodes an RNA-dependent RNA polymerase (RdRp) and is proposed to confer resistance to TYLCV by amplifying the RNAi signal  viruses. The major class of these genes represent single dominant resistance genes (and of which the biggest group consists of the NB-LRR type), while others are recessive, tolerance, or partial resistance genes. A very nice example of a dominant resistance gene of the latter case has recently been described with the cloning and characterization of the S genes) are proposed to provide a more durable resistance than dominant R genes. However due to their functions they may cause pleiotropic effects when knocked out from the host genome  Figure : a stratR) gene products (in)directly sense the presence of a specific pathogen by their effector, termed avirulence factors (Avr), as a counter defense against ETS, leading to a stage called Effector-Triggered Immunity (ETI)  induction of a programmed cell death response, as visualized by the rapid appearance of necrotic lesions  or in rare occasions extreme resistance (ER) during which no necrosis is observed at all. However, more and more evidence is presented, that there is an uncoupling of the resistance response from the programmed cell death response, although both can work in concert. Due to these responses, viruses (and other pathogens) are confined to the site of entry/invasion where infections are prevented. In contrast to the slower onset of antiviral RNAi, the R gene response generally is rapid and within ~3/4 days lead to containment of the virus.Single dominant resistance  Figure . TriggerR genes basically can be grouped into two classes, namely those that encode NB-LRRs and all others. The major class of R genes consists of the NB-LRR type and encode proteins that, irrespective of the pathogen they recognize, consist of three domains; (1) the Nucleotide Binding Site (NBS) in the center of the protein, (2) a Leucine Rich Repeat (LRR) at the C-terminal end, and (3) a Coiled-coil (CC) or Toll and Interleukin-1 Receptor (TIR) domain at the N-terminal end of the resistance gene product  , only 22 have been cloned and characterized. Some R genes have functional alleles in other plant species, often showing a similar Avr recognition. The majority of the known R gene products are of the CC-NB-LRR type, whereas only a small group belongs to the TIR-NB-LRR group , as commonly observed with NB-LRR mediated resistance responses  and thereby inhibits RNA replication  presents a housekeeping gene  or clearly act as homologs (Rx1 and Rx2 from S. tuberosum) and recognize the same Avr protein from overlapping virus species, indicating that these conserved R proteins are able to recognize similar structures but with an adapted spectrum , the movement protein , the replicase protein  or the RNAi suppressor protein , all potentially can act as elicitor of resistance  R genes to a wide range of various pathogens; e.g., Rx1 and GPa2 both interact with the same guardee RanGAP2  and activates a downstream signaling pathway leading to resistance , an HR is induced in N. tabacum, a response that does not occur in N. benthamiana. Bhattacharjee et al. . As a result, synthesis of viral proteins is inhibited, ultimately preventing virus accumulation and spread. Whether this mechanism is generic to all R genes against plant-viruses remains to be investigated.The downstream mechanism after resistance induction still remains unclear. However, one described case of the e et al.  employedR genes is tightly controlled through the activity of miRNAs. One study showed the miR482/2118 superfamily negatively controlled the translation of NB-LRR proteins by targeting its P-loop motif  required.More recently, two independent studies were published that showed that the translation of R genes trigger a hypersensitive response (HR) or an extreme response (ER) in case the reaction occurs in a single cell. Both involve a programmed cell death (PCD) response that rapidly kills infected cells and prevents systemic spread of the (virus) pathogen. An induced HR is quite characteristic and involves the activation and expression of SA, jasmonic acid (JA), nitride oxide (NO), ethylene, reactive oxygen species (ROS), and Ca2+, and expression of Pathogenesis Related (PR)-genes. While each component has a specificity toward certain pathogens, only SA, ROS, and Ca2+ seem to be effective against viruses  and resistance are distinct physiological pathways  , more recently it has been found that there are also proteins that modulate the strength of defense responses   share similarity to these R genes, however both function as PRRs in the PTI response. The first major class present the “nucleotide-binding domain and leucine-rich repeat”-proteins (NLRs) and the second class is that of Toll like receptors (TLRs), which are all found to function as PRRs in the PTI response. Both are immune receptors aimed at detecting “foreign” structures and activating downstream defense responses. The family of NLRs share the most homology, as evidenced when looking at R genes from plants and NACHT-LRR encoding genes from the animal kingdom (NAIP–CIITA–HET-E–TP1 domain)   in these proteins. The most prevalent type of R proteins in plants belong to the NB-LRR protein structural class, from which the central nuclear binding domain (NBS) exhibits similarity to the nucleotide binding domain in several metazoan apoptosis regulating proteins like Apaf-1 from mammals and CED-4 from TLRs represent the best studied family of PRRs in mammals so far. They are transmembrane glycoprotein receptors with an extracellular PAMP-binding domain consisting of multiple LRR that fold into a “horseshoe” structure. Additionally, it possesses intracellular signaling regions that have similarity to the intracellular domain of the Interleukin-I receptor ((IL-1R), which is referred to as Toll/IL-1R (TIR) domain that mediates downstream signaling upon activation of the receptor. TLRs initiate signal cascades involving the activation of nuclear factor kappa b (NF-κ B), mitogen-activated protein kinase (MAPK), and interferon regulatory factors (IRFs). This subsequently leads to a concerted expression of interferons, cytokines, and chemokines. Finally, inflammatory processes, cell cycle arrest, and cell death are induced (Honda et al., Pathogen recognition in both animal and plant kingdoms involves the LRR domain, which binds the ligand in its horseshoe shape, often followed by activation of a signaling cascade through kinase phosphorylation. Structural similarities between animal TLR/NLR and plant NB-LRR proteins point to a convergent evolution of these defense-related pathways (Yue et al., R gene mediated defense and the induction of the HR. However, the number of well-studied examples is still very limited, and therefore makes it difficult to extrapolate to other less studied R genes. In depth studies on other R genes from different crops providing resistance against different phytopathogens will have to show whether there is common mechanism of defense shared between all R genes, or whether specialization between different classes of R genes occurs. Knowledge on mammalian innate immunity sensors and their mode of action may provide interesting and cross pollinating views in this. The rapid generation of resistance breaking virus isolates against dominant resistance genes already indicates the importance for alternative resistance genes, to provide more durable, and effective resistances.Dominant resistance against plant viruses are of increasing interest to breeders and scientists in the past years. More and more is known about the molecular mechanisms behind The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The recent increase in the use of high field MR systems is accompanied by a demand for acquisition techniques and coil systems that can take advantage of increased power and accuracy without being susceptible to increased noise. Physical location and anatomical complexity of targeted regions must be considered when attempting to image deeper structures with small nuclei and/or complex cytoarchitechtonics , such as the brainstem and the cerebellum (Cb). Once these obstacles are overcome, the concomitant increase in signal strength at higher field strength should allow for faster acquisition of MR images. Here we show that it is technically feasible to quickly and accurately detect blood oxygen level dependent (BOLD) signal changes and obtain anatomical images of Cb at high spatial resolutions in individual subjects at 7 Tesla in a single one-hour session. Images were obtained using two high-density multi-element surface coils  placed beneath the head at the level of Cb, two channel transmission, and three-dimensional sensitivity encoded  acquisitions to investigate sensorimotor activations in Cb. Two classic sensorimotor tasks were used to detect Cb activations. BOLD signal changes during motor activity resulted in concentrated clusters of activity within the Cb lobules associated with each task, observed consistently and independently in each subject: Oculomotor vermis (VI/VII) and CrusI/II for pro- and anti-saccades; ipsilateral hemispheres IV-VI for finger tapping; and topographical separation of eye- and hand- activations in hemispheres VI and VIIb/VIII. Though fast temporal resolution was not attempted here, these functional patches of highly specific BOLD signal changes may reflect small-scale shunting of blood in the microvasculature of Cb. The observed improvements in acquisition time and signal detection are ideal for individualized investigations such as differentiation of functional zones prior to surgery. Cb was isolated from the T1w-image using the SUIT toolbox . Note thSignal quality of these surface coils has been previously defined for visual cortex ,55 showip < 0.05 (voxel-based family-wise error (FWE) false discovery rate (FDR) corrected, automatically calculated with SPM interface, further referred to as ‘SPMT’) frequently occurred in extremely small clusters that are difficult to make out at any scale without significant prior knowledge of Cb anatomy. This default FDR is a very strict correction for individual analyses and does not weigh the individual variations in noise. Therefore a cluster-based FDR correction utilizing the intercepts of the noise Gaussian distribution with the positive and negative gamma signal was also applied, specified for each subject individually , and a high-pass filter of 80s was applied to each timeseries; this model also included the aforementioned WM and CSF nuisance regressors. Timeseries from all voxels within the imaged volume were included when estimating the model. The resulting T-maps from this active versus rest contrast were overlaid on the T1w images using MRIcron . Voxels ds, ‘AT’ ). The ATTo assess the stability of the results across the sample size of this experiment, statistical maps from each subject were created from the SUIT-normalized and smoothed functional images masked inclusively for voxels within the standard SUIT Cb mask. All resulting maps were overlaid on the SUIT atlas to investigate the validity of the individual activations in common (MNI) space. These maps were then entered into two simple second level analyses (Paired t-Test and multiple regression only) in SPM to evaluate activations at the group level. No additional second level analyses were run, as the goal of this study was to investigate activations at the single-subject level.The data of one subject were acquired when the Cb was below the FOV of the coils and these data are not displayed in the figures of the main text; see p < 0.05 thresholds for the two types of FDR thresholding analyses run: from cluster-based AT FDR values  to voxel-based family-wise error (FWE) SPMT FDR values . In Figs Reliable signal changes were observed within Cb when contrasting active versus rest periods for both tasks in all subjects. Figs vVI-VII) and hVI) where activity was indeed observed in each subject during PA eye movements. Note that activations were confined to the grey matter and that the pinpoint PA-related activity is located in clusters bilaterally along lobule VI in the axial plane , seen as bright yellow activations in line with the curvature of the lobule marked by crosshairs in the axial panels of hV where finger-movement related activity was also observed as expected and reliably in each subject. Also note the restriction of FT-related activity to a small number of folia within ipsilateral (R) hV-VI (Crosshairs in R) hV-VI . AlthougR) hV-VI , and maxR) hV-VI hCrusI and/or hCrusII as well as hVIIb during the PA task  afferents from Cb cortex to ipsilateral dentate nucleus (D) mean that activations in the PC layer (middle Cb cortex layer) should result in deactivation of D. The conspicuity of structural detail in the T1w scan is apparent, with remarkable definition of the folia, especially around the edges of the Cb Figs –7 where left) and right hV for FT-related activity displayed in right). Select cross sections of Cb are shown in 8b and 8c to display the variable location of activity between individuals within these lobules.The activation loci and T values for activations in each lobule of interest  are visible on the top of each panel and just below the axial slice, respectively, for each subject Figs –6. DespihVI . In addition, some activity was normalized outside of Cb, and inter-subject activations infrequently overlap within a single lobule. The remarkable differences in individual functional organization remain in both individual and common MNI space analyses. For example S01 and S06 infrequently overlap with any other subject, while S03 and S05 show a lot of PA overlap and S02 and S03 show a lot of FT overlap with each other in common space  was located caudal to that related to motor (FT) activity  acquisitions to image subsystem-related activity in Cb in individual subjects. We were able to clearly differentiate activity throughout the entire Cb cortex including D in the DCN in a single session lasting less than one hour per subject. BOLD signal changes during motor activity resulted in concentrated clusters of activation observed within the expected lobules for each task in each subject: OMV and activity  as seen Patients with Cb disorders often show abnormalities in eye movements and finger tapping (and/or other motor dysfunctions) ,24, behahVI and hCrusII/hVIIb activations during PA are consistently distinct from ipsilateral hV-VI and hVIII activations during FT  is also possible within  seen by ; though It is possible that inter-subject differences in Cb physiology and/or arrangement of functional patches underlie the lack of overlap in normalized space; or this could be a result of the normalization method. Current normalization schemes ,67 that h, L paravermis, vermis, R paravermis, Rh) and are arguably most significant outside of OMV; visible in the axial slices of hVI in response to attending a moving grating [hCrusI in response to saccadic errors [hV-VI, OMV, and throughout CrusI/II in response to attended flashed visual stimuli [or finger-movement related, suggesting that observed patches of activity are related to motor planning and output rather than visual perception or attention alone since subjects were also attending these stimuli during the FT task. However, the location of these dots was only relevant during the PA task. It is therefore possible that the observed PA-related clusters, acting alone or in concert with CrusI/II, represent either possible task-relevant attention, target locations  or target awareness, or a mapping of visual space in order to monitor and/or update performance on a visual task. Although the presence of vector mapping of saccades within Cb is currently unknown, spatial saccade maps are known to exist in superior colliculus and frontal eye fields [The clustered PA-related activations are located in five bilateral patches along lobule VI , despite the fact that these high-density multi-element coils only cover a part of the head. Since signal CNR was sufficient to consistently differentiate eye-related from hand-related motor activity and to confirm suspected cognition-related activity  in each subject scanned, we can conclude that these coils are beneficial for fast imaging of Cb. Furthermore, the use of localized shimming and 3D-EPI acquisitions successfully avoided image distortion, artifacts, and signal losses commonly observed around Cb at high field strength and the entire scanning session lasted less than one hour.Combining this acquisition scheme with newer analysis methodologies can allow for a larger number of investigations within one individual in a single scan session while freeing up space within the transmit coil. Future studies targeting event-related perception and learning paradigms can utilize these methods, specifically in human and non-human primate Cb, to further elucidate the neurovascular interplay of superficial and deep Cb and the dynamics and organization of zones or patches in humans to close the gap of inference with regard to the affects of neuronal processes on BOLD signal changes.S1 FigMethods: Coil Comparison Signal quality of the surface coils has been previously defined for visual cortex [Figure: Coil Comparison—PA Functional Activations.Bottom: OMV , Top: hVI, and CrusII  are activated during the PA task—activation are shown in red-to-yellow, Left: Activations observed using a 32-channel full head coil  Right: Activations observed using two 16-channel surface coils with the same subject. CrusII activations are also observed with the head coils (See l cortex  but not (TIF)Click here for additional data file.S2 FigExample images from one subject where coils were placed dorsal to Cb. Although most activation is limited to dorsal Cb, some activity in ventral Cb is still observable. Images can be compared to activations from the other six subjects with correct coil placement. Slice locations  are displayed at the top of each panel and T-values at the crosshairs are displayed at the bottom of the axial (bottom left) panels. Refer to (TIF)Click here for additional data file.S3 FighVIIb and R hCrusII), indicated by crosshairs. It is clear that significant changes in BOLD signal are not restricted to areas with higher tSNR (yellow-white regions of tSNR images) nor do they occur spuriously around Cb vasculature (dark dots in the T2*w images). Slice locations  are displayed at the top of each Funct panel and coordinates are the same for all three images. T-values and tSNR strength at the crosshairs are displayed at the bottom of the axial (rightmost) panels. Refer to T maps (Funct) and tSNR from a single PA run are overlaid on the T1w structural scan from one subject and aligned with the T2*w (T2*) structural images for six Cb ROIs Click here for additional data file.S1 TableNM—Normalised and Smoothed. *Thumb was tapped during the ENTIRE session (no rest period), so FT was not compared between head and surface coils.(PDF)Click here for additional data file.


Patients with chronic obstructive pulmonary disease (COPD) suffer from significantly more cardiovascular comorbidity and mortality than would be anticipated from conventional risk factors. The aim of this study was to determine whether COPD patients have a higher coronary artery calcium score (CACS) and epicardial fat burden, compared to control subjects, and their association with cardiovascular events.From a registry of 1906 patients 81 patients with clinically diagnosed COPD were one-to-one matched to 81 non-COPD control subjects with a smoking history, according to their age, sex, and the number of classic cardiovascular risk factors . CACS, epicardial fat, and subsequent major adverse cardiovascular events (MACE) during follow-up were compared between groups.3 [95% confidence interval -20.9 to 21.9], p=0.961) compared with controls. After a median follow-up of 42.6 months a higher incidence of MACE was observed in COPD patients  compared with controls. Cox proportional hazard regression identified cardiac ischemias and CACS as independent predictors for MACE.Patients with COPD  showed no difference in CACS  or epicardial fat volume (mean difference -0.5 cmCOPD patients experienced a higher MACE incidence compared to controls despite no baseline differences in coronary calcification and epicardial fat burden. Other mechanisms such as undersupply of medication seem to account for an excess cardiovascular comorbidity in COPD patients. Chronic obstructive pulmonary disease (COPD) is characterized by irreversible airflow limitation. Globally, COPD is the fourth leading cause of death in industrialized countries. The clinSingle-photon emission computed tomography (SPECT) is a robust diagnostic imaging tool which can assess previously unknown myocardial scar tissue or ischemia as potential predictors for future risk assessment. FurthermThere are few and conflicting data on the relationship between COPD and subclinical CAD and some of the published studies are limited by inadequate matching of classic cardiovascular risk factors between COPD patients and control subjects.–12 The e2). MACE were defined as documented death due to a cardiovascular event, non-fatal infarction (including stroke), documented transient ischemic attack, catheter-based target lesion revascularization, unstable angina pectoris requiring hospitalization, and coronary artery bypass graft.In this one-to-one matched cohort study COPD patients  were screened for the following inclusion criteria: (1.) coronary artery calcium score (CACS) data available; (2.) history of smoking (smoker or ex-smoker with a smoking history of ≥10 pack-years [PY]); (3.) objectively confirmed COPD. COPD was staged according to Global Initiative for Chronic Obstructive Lung Disease [GOLD]-guidelines 2007 and spirEach COPD subject was 1-to-1 matched with one non-COPD subject for: (1.) sex (exact), (2.) number of classic cardiovascular risk factors (exact), and (3.) age. Age  was matched via nearest neighbour matching with a maximum allowable difference of one standard deviation between two participants. Matching was performed without replacement and optimal in order to minimize the total within pair differences. COPD subjects for whom no control case was found within the maximum absolute distance (n = 25) were left unmatched and excluded from analysis. Hence, only 81 (76%) of the COPD subjects were included in the analysis. Balance assessment in measured baseline covariates was additionally performed with the standardized difference, which is the difference in means in units of the pooled standard deviation.This study complies with the Declaration of Helsinki of the World Medical Association, ICH-GCP-Guidelines. The study protocol was approved by the Local Ethics Committee of the Canton of Zurich (KEK-ZH-Nr. 2013–0615) and registered with the study ID NCT02162095 . Patients were included in the study if they had signed informed consent authorizing their records to be included in our cardiac imaging research registry. All patients underwent clinically indicated cardiac SPECT. Informed consent for the longitudinal part of the study (follow-up) was obtained separately from all patients or relatives in case of a deceased patient which was recorded by an investigator on the case report form. The ethics committee approved this procedure due to the low risk category this study entails.All patients underwent a one-day pharmacological stress/rest SPECT Myocardial Perfusion Imaging protocol according to the guidelines of the European Association of Nuclear Medicine. All card3. Epicardial and thoracic fat were cranially confined by the first CT-slice displaying the bifurcation of the pulmonary trunk and caudally by the most inferior CT-slice of the myocardium. Epicardial fat was defined as detected fat-voxels within the visceral pericardium including all fat surrounding the coronary arteries while thoracic fat was defined as detected fat-voxels within the thoracic cavity limited by the posterior limit of the heart.[The CACS was calculated in clinical routine using a commercially available semi-automatic software  according to the Agatston scoring method and expressed in Agatston Units (AU). The areahe heart. Thereforhe heart. The thorhe heart.Follow-up was conducted either via personal contact, or with the help of the Swiss Federal Department of Justice and Police (Swiss register “Infostar”). Information was cross-checked and validated from internal and requested external medical charts with formal approval of the patient. All medical charts were obtained and MACE were critically reviewed by two investigators  unaware of the participants allocation. All deaths were categorized into cardiac or non-cardiac. The freedom from MACE time of a patient started at the time of SPECT imaging and ended at death/MACE, or at the last follow-up day. This method did not account for recurring MACE in one subject. Therefore, overall MACE rate was expressed as MACE/follow-up year.Continuous data are expressed as mean (±SD), or in the case of non-normal distribution by their median (quartiles). Paired tests (shared frailty) were used for matched pairs in this study. Ln(CACS+1) was used to investigate CACS-relationships with other variables. Continuous and categorical baseline characteristics of COPD and non-COPD subjects were compared using Wilcoxon and Pearson chi-square tests respectively. Spearman correlation analysis was used to investigate correlations with the CACS. Cumulative freedom from MACE at follow-up was analysed by means of the Kaplan—Meier method and compared between groups using the log-rank test. Univariable and multivariable (stepwise backward selection manner) Cox proportional hazard regression models were used to identify possible predictors of MACE. Furthermore, age, sex, and number of cardiovascular risk factors were also adjusted for multivariable analysis. Data analysis was conducted using IBM SPSS Statistics v. 22.0 software . A two-sided p value of <0.05 was considered to indicate statistical significance.Of the 1906 patients screened, 81 matched pairs entered the data analysis. The study population had a mean age of 64 years and was predominantly male (78%). All COPD GOLD stages were represented, but most of the COPD patients were in GOLD stage IV . Apart from the anticipated differences in the lung function test and consumed PY, the two cohorts differed in BMI (but not obesity) and number of prescribed ß-blockers. Follow-up was successful in 154 (95%) patients, five patients were lost in the COPD group and three patients in the control group.3, p = 0.961 and 277±139.5 vs. 283.0±149.8 cm3, p = 0.931 respectively). Epicardial and thoracic fat correlated highly  whereas correlation of epicardial and thoracic fat with CACS  and BMI  was moderate. CACS and epicardial fat correlated with the number of cardiovascular risk factors and detected scars in the SPECT .CACS did not differ significantly between the COPD group and the control group  nor in any GOLD stage compared to the controls adjusted for age, sex, number of cardiovascular risk factors, and smoking status (data not shown). Fat-quantification was successful in all but one patient . There was no significant mean (SD) group difference in epicardial and thoracic fat between the COPD group and the control group  and 48 deaths  occurred. Median follow-up was 42.6  months and did not differ between groups (p = 0.543). Kaplan-Meier analysis  indicate1 (% pred.), PY, ischemia in SPECT imaging, and previous CAD were risk factors for MACE  there is no therapy which has a profound effect on disease severity. The relatively small sample size mainly due to restrictive 1-to-1 matching and limited amount of COPD patients undergoing SPECT also remains a limitation of this study.In conclusion, there is an excess cardiovascular comorbidity in COPD patients which cannot be explained by coronary calcification or epicardial fat burden and thus further well designed studies with a focus on medication and comorbidities are needed to investigate the mechanisms underpinning the association between COPD and cardiovascular disease.


Many immunohistochemical markers have been used in the postmortem detection of early myocardial infarction.In the present study we examined the role of Heart-type fatty acid binding protein (H-FABP), in the detection of early myocardial infarction.We obtained samples from 40 human autopsy hearts with/without histopathological signs of ischemia.All cases of definite and probable myocardial infarction showed a well-defined area of H-FABP depletion. All of the control cases showed strong H-FABP expression, except two markedly autolysed myocardial samples that showed affected antigenicity.Thus, we suggest H-FABP as being one of the valuable tools facing the problem of postmortem detection of early myocardial infarction/ischemia, but not in autolysis. Myocardial infarction (MI) is the most frequent diagnosis made in the majority of sudden deaths subjected to clinical and medico legal autopsies . The proIt is impossible to get the histological diagnosis of early myocardial infarction as a cause of sudden death in routine H&E stained sections, as it is unlikely to find special changes for myocardial infarction aged less than 6 hours , 4.Thus, in the current autopsy practice, there is a great need for more sensitive diagnostic methods for the postmortem diagnosis of early myocardial damage .Also, we need further studies on the postmortem stability of immunohistochemical markers and the regularity of their expression alteration in different postmortem intervals to define the role of autolysis .Heart-type fatty acid binding protein (H-FABP) is a low molecular weight protein (14-15kDa), which is abundant in the cytoplasm of myocardial cells. As myocardial cell membrane is damaged by ischemia, H-FABP leaks to the extracellular space and enters the blood circulation very easily and quickly due to its small size and water solubility .Therefore, loss of H-FABP in myocardial ischemic cells could occur as early as 15 minutes, and as the ischemia intervals prolonged, the depletion areas increase gradually in myocardial cells. The deletion areas of H-FABP reach a peak after four hours of myocardial ischemia .The primary results suggest that H-FABP staining can detect very early ischemic damages in human myocardium , 9.In the present study we examined the role of Heart-type fatty acid binding protein (H-FABP), in the detection of early myocardial infarction.The study included 40 hearts from autopsy cases; with/without evidence of coronary artery disease (CAD). CAD cases included advanced or complicated lesions.Tissue sections, from the myocardia and the corresponding coronary artery supply were obtained. They were fixed in 10% neutral-buffered formalin and embedded in paraffin. H&E stained sections are obtained and examined histologically.Cases were grouped according to the histological findings as GI, GII, and GIII according to the Lodge Patch histological criteria for MI :Group I: includes 15 hearts represented cases showing signs of old ischaemia .Group II: includes 18 cases represented cases showing either signs of recent ischemia  or with evidence of advanced or complicated CAD .Group III: includes 7 cases showing no signs of ischemia, two of them showed severe autolytic changes which were selected to assess the expression of H-FABP in autolysis.Death/Autopsy time ranged between 12-24 hours in all examined cases, except for the two autolysed control cases where death/autopsy time exceeded 60 hours.Finally, we obtained 40 sections of myocardial tissue from the 40 cases for immunohistochemical staining by H-FABP.Immunohistochemical staining with H-FAB is performed using a standard avidin–biotin–peroxidase system.Approximately 4 µm thick histologic sections were deparaffinated in xylene and alcohol, rehydrated in distilled water for 5 minutes, and then washed in PBS for 5 minutes. To reveal the antigens, they were pre-treated by using the proteolytic enzyme proteinase K and then washed in PBS for 5 minutes.The primary monoclonal antibody to H-FABP  was diluted to a concentration of the antibody 1:250  and incubated for 60 minutes at 37°C, then washed in PBS for 5 minutes.Secondary antibody was applied for 60 minutes , followed by rinsing with PBS. The reaction was visualized with DAB chromogen . The slides were counterstained with diluted hematoxylin.Each slide has its built in negative and positive controls for H-FABP. Areas with definitive signs of ischaemia by hematoxylin and eosin (H&E) acted as negative internal control, whereas non- ischemic regions of myocardium are expected to be positive internal control.The amount and extent of depletion of the cell marker is recorded. Loss of myocardial staining of H-FABP in the three groups was recorded. The obtained results were scored from (0) to (-3) according to the percent of myocardial fibers showing depletion of H-FABP staining: Score 0: no loss of H-FABP staining, Score -1: loss in less than 25% of fibers, Score -2: loss in 25-50% of fibers, Score -3: loss in >50% of fibers .The results are summarized in the Myocardial ischemia with Hematoxylin and eosin stain (x 200) is shown in Group I ; all casGroup II Figures  and 1D; Group III; 5 of the selected 7 cases showed strong brown cytoplasmic staining with no loss (score 0), denoting the integrity of the myocardium . The remHistopathological examination of the heart must be performed as a part of routine autopsy as to clarify the cause of death, especially in non-traumatic deaths in which the autopsy didn’t reveal any confessant non-cardiac cause of death .Many studies have proved the usefulness of variable immunohistochemical markers  in the diagnosis of early myocardial injury has been promoted because most of them can be visible very early after the beginning of the symptoms (chest pain and angina attacks) -16.However, very few studies have indicated the usefulness of H-FABP in cardiology for detection of acute myocardial infarction (AMI) in both human and animal models , 8.On studying H-FABP immunostaining, all group I cases with old myocardial infarction, showed H-FABP depletion in the infarcted areas.These results are consistent with the results obtained by Meng et al., 2006, who recognized H-FABP depletion in infarcted area in the human myocardia . They arWe also noticed variable degrees of H-FABP depletion in the myocardial fibers surrounding the infarcted areas that appeared normal in the H&E stained slides. This was rather more remarkable than the results of Ming et al., 2004 who declared that these apparently normal cardiomyocytes were brown mixed with some faintly immunostained cardiomyocytes .An example from this group was a heart with left ventricular concentric hypertrophy. Microscopic examination revealed severe stenosis in his left circumflex artery and minimal foci of myocardial fibrosis in the lateral wall of left ventricle. H-FABP staining of the section selected from and around the fibrotic foci revealed wider areas of ischemia - loss (-2) - around fibrotic foci denoting more pronounced ischemia, which could only be detected by the antibody, giving the impression of its high sensitivity.Selected group II cases, with probable signs of ischaemia had coronary stenosis and their complication showed loss of cytoplasmic staining by various degrees ranging from minimal depletion (-1) and up to near total depletion (-3). Meng et al, 2006, in their study, noticed H-FABP depletion in areas of suspected early myocardial infarction that showed normal H&E staining .th decade of life. He died in custody half an hour after exposure to a homicidal attempt by strangulation by another prisoner. On autopsy, there were only bruises on neck side. Examination of the heart revealed no gross myocardial abnormalities. Microscopic examination revealed severe stenosis of the left anterior descending coronary artery with rupture in arterial wall atheromatous plaque complicated by recent thrombus formation. Myocardium showed only congestion and an area of fragmentation by H&E staining. However; immunohistochemical examination of the myocardium revealed areas of H-FABP depletion (score -2) documenting early ischemic changes.Worth mentioning, one of group II cases belonging to a male in his 5Acute coronary syndrome found in this case led to death with the subsequent myocardial ischemia only detected by Immunohistochemical staining .On examining group III cases - control positive group- five out of the seven cases showed diffuse cytoplasmic H-FABP immunoreactivity exhibited in the cardiac myocytes (score 0). In comparison with the study performed by Meng et al., 2006, cases with no signs of ischemia, showed weak, but diffuse staining .The remaining two cases that showed depletion in this group were two autolysed hearts from two putrefied female cadavers, with no evidence of ischemia. Estimated time of death exceeded 60 hours. Both cases showed H-FABP depletion, which reflects that H-FABP immunoreactivity in this study was markedly affected by autolysis.This is contradictory to the results recorded by Ming and Meng, 2004. They stated that the efficacy of H-FABP for detection of ischemic cardiomyocyte lesions was not reduced by autolysis, even in cases where death autopsy time extended as long as 60 hours after death on human hearts .We can summarize that H-FABP may be a useful marker in the early detection of AMI as well as the ongoing ischemia in areas surrounding well established infarctions in human hearts which can’t be detected using routine H&E stain, as well as the ongoing ischemia in areas surrounding well established infarctions.However; it is not the marker of choice that can be used in the detection of early myocardial ischemia in advanced autolytic changes - despite its sensitivity generally. Other antibodies that can withstand autolysis as troponin, myoglobin and desmin can do that role instead , 11, 17.Many factors are needed for well-established confident diagnosis besides the immunohisto-chemistry. We should correlate with the history, clinical data and case circumstances and their insufficiency may act as an obstacle in achieving satisfactory results that we are aiming at. In forensic practice, we can’t give opinion depending on single factor, there are many factors should be put together to get out at the end by reliable confident decision, and every case in our practice has its unique characteristics, circumstances and findings that make it an indivisible standalone entity.