Disentangling the Role of Cortico-Basal Ganglia Loops in Top-Down and Bottom-Up Visual Attention: An Investigation of Attention Deficits in Parkinson Disease.

It is solidly established that top-down (goal-driven) and bottom-up (stimulus-driven) attention mechanisms depend on distributed cortical networks, including prefrontal and frontoparietal regions. On the other hand, it is less clear whether the BG also contribute to one or the other of these mechanisms, or to both. The current study was principally undertaken to clarify this issue. Parkinson disease (PD), a neurodegenerative disorder primarily affecting the BG, has proven to be an effective model for investigating the contribution of the BG to different brain functions; therefore, we set out to investigate deficits of top-down and bottom-up attention in a selected cohort of PD patients. With this objective in mind, we compared the performance on three computerized tasks of two groups of 12 parkinsonian patients (assessed without any treatment), one otherwise pharmacologically treated and the other also surgically treated, with that of a group of controls. The main behavioral tool for our study was an attentional capture task, which enabled us to tap the competition between top-down and bottom-up mechanisms of visual attention. This task was suitably combined with a choice RT and a simple RT task to isolate any specific deficit of attention from deficits in motor response selection and initiation. In the two groups of patients, we found an equivalent increase of attentional capture but also comparable delays in target selection in the absence of any salient distractor (reflecting impaired top-down mechanisms) and movement initiation compared with controls. In contrast, motor response selection processes appeared to be prolonged only in the operated patients. Our results confirm that the BG are involved in both motor and cognitive domains. Specifically, damage to the BG, as it occurs in PD, leads to a distinct deficit of top-down control of visual attention, and this can account, albeit indirectly, for the enhancement of attentional capture, reflecting weakened ability of top-down mechanisms to antagonize bottom-up control.

Time-frequency modulation of ERD and EEG coherence in robot-assisted hand performance.

A better understanding of cortical modifications related to movement preparation and execution after robot-assisted training could aid in refining rehabilitation therapy protocols for stroke patients. Electroencephalography (EEG) modifications of cortical activity in healthy subjects were evaluated using time-frequency event-related EEG and task-related coherence (TRCoh). Twenty-one channel EEG was recorded in eight subjects during protocols of active, passive, and imagined movements. The subjects performed robot-assisted tasks using the Bi-Manu-Track robot-assisted arm trainer. We applied time-frequency event-related synchronization/desynchronization (ERS/ERD) and TRCoh approaches to investigate where movement-related decreases in power were localized and to study the functional relationships between areas. Our results showed ERD of sensorimotor (SM) area over the contralateral side before the movement and bilateral ERD during execution of the movement. ERD during passive movements was similar in topography to that observed during voluntary movements, but without pre-movement components. No significant difference in time course ERD was observed among the three types of movement over the two SM areas. The TRCoh topography was similar for active and imagined movement; before passive movement, the frontal regions were uncoupled from the SM regions and did not contribute to task performance. This study suggests new perspectives for the evaluation of brain oscillatory activity and the neurological assessment of motor performance by means of quantitative EEG to better understand the planning and execution of movement.

Spatial and Temporal EEG-fMRI Changes During Preictal and Postictal Phases in a Patient With Posttraumatic Epilepsy.

The combined use of electroencephalography (EEG) and functional magnetic resonance imaging (EEG-fMRI) in epilepsy allows the noninvasive hemodynamic characterization of epileptic discharge-related neuronal activations. The aim of this study was to investigate pathophysiologic mechanisms underlying epileptic activity by exploring the spatial and temporal distribution of fMRI signal modifications during seizure in a single patient with posttraumatic epilepsy. EEG and fMRI data were acquired during two scanning sessions: a spontaneous critical episode was observed during the first, and interictal events were recorded during the second. The EEG-fMRI data were analyzed using the general linear model (GLM). Blood oxygenation level-dependent (BOLD) localization derived from the preictal and artifact-free postictal phase was concordant with the BOLD localization of the interictal epileptiform discharges identified in the second session, pointing to a left perilesional mesiofrontal area. Of note, BOLD signal modifications were already visible several seconds before seizure onset. In brief, BOLD activations from the preictal, postictal, and interictal epileptiform discharge analysis appear to be concordant with the clinically driven localization hypothesis, whereas a widespread network of activations is detected during the ictal phase in a partial seizure.

Investigation of brain hemodynamic changes induced by active and passive movements: a combined arterial spin labeling-BOLD fMRI study.

PURPOSE: To assess the applicability of arterial spin labeling (ASL) in comparison to blood-oxygenation-level-dependent (BOLD) contrast fMRI in detecting brain activations elicited by active and passive hand movements. MATERIALS AND METHODS: A block design for ASL and BOLD fMRI was applied in 8 healthy subjects using active and passive hand tasks. Data analyses were performed at individual and group level, comparing both the different movements and the performance of the two techniques. RESULTS: Group analyses showed involvement of the same areas during both tasks, as the contralateral sensorimotor cortex, supplementary motor area, cerebellum, inferior parietal lobes, thalamus. ASL detected smaller activation volumes than BOLD, but the areas had a high degree of colocalization. Few significant differences (P < 0.05) were found when the two tasks were compared for the number of activated voxels, coordinates of center of mass, and CBF estimates. Considering together all the areas, the mean %BOLD change was 0.79 ± 0.27 and 0.73 ± 0.24 for the active and passive movements respectively, while the mean %CBF changes were 34.1 ± 8.9 and 27.1 ± 14.8. CONCLUSION: Our findings confirm passive and active tasks are strongly coupled, supporting the importance of passive tasks as a diagnostic tool in the clinical setting. ASL fMRI proved suitable for functional mapping and quantifying CBF changes, making it a promising technique for patient cohort applications.

The cerebellum and visual perceptual learning: evidence from a motion extrapolation task.

Visual perceptual learning is widely assumed to reflect plastic changes occurring along the cerebro-cortical visual pathways, including at the earliest stages of processing, though increasing evidence indicates that higher-level brain areas are also involved. Here we addressed the possibility that the cerebellum plays an important role in visual perceptual learning. Within the realm of motor control, the cerebellum supports learning of new skills and recalibration of motor commands when movement execution is consistently perturbed (adaptation). Growing evidence indicates that the cerebellum is also involved in cognition and mediates forms of cognitive learning. Therefore, the obvious question arises whether the cerebellum might play a similar role in learning and adaptation within the perceptual domain. We explored a possible deficit in visual perceptual learning (and adaptation) in patients with cerebellar damage using variants of a novel motion extrapolation, psychophysical paradigm. Compared to their age- and gender-matched controls, patients with focal damage to the posterior (but not the anterior) cerebellum showed strongly diminished learning, in terms of both rate and amount of improvement over time. Consistent with a double-dissociation pattern, patients with focal damage to the anterior cerebellum instead showed more severe clinical motor deficits, indicative of a distinct role of the anterior cerebellum in the motor domain. The collected evidence demonstrates that a pure form of slow-incremental visual perceptual learning is crucially dependent on the intact cerebellum, bearing the notion that the human cerebellum acts as a learning device for motor, cognitive and perceptual functions. We interpret the deficit in terms of an inability to fine-tune predictive models of the incoming flow of visual perceptual input over time. Moreover, our results suggest a strong dissociation between the role of different portions of the cerebellum in motor versus non-motor functions, with only the posterior lobe being responsible for learning in the perceptual domain.

Paroxysmal dysarthria-ataxia in remitting-relapsing Bickerstaff's-like encephalitis.

Paroxysmal dysarthria-ataxia is a rare neurological condition due to ephaptic transmission, generally appearing in multiple sclerosis patients characterized by stereotyped attacks of slurred speech usually accompanied by ataxia, appearing many times a day. Here we describe a patient with an unusual remitting-relapsing form of Bickerstaff's-like brainstem encephalitis who manifested PDA after a relapse with the involvement of a peculiar region below the red nuclei and benefited from lamotrigine.

EEG-fMRI evaluation of patients with mesial temporal lobe sclerosis.

This preliminary study sought more information on blood oxygen level dependent (BOLD) activation, especially contralateral temporal/extratemporal spread, during continuous EEG-fMRI recordings in four patients with mesial temporal sclerosis (MTS). In two patients, EEG showed unilateral focal activity during the EEG-fMRI session concordant with the interictal focus previously identified with standard and video-poly EEG. In the other two patients EEG demonstrated a contralateral diffusion of the irritative focus. In the third patient (with the most drug-resistant form and also extratemporal clinical signs), there was an extratemporal diffusion over frontal regions, ipsilateral to the irritative focus. fMRI analysis confirmed a single activation in the mesial temporal region in two patients whose EEG showed unilateral focal activity, while it demonstrated a bilateral activation in the mesial temporal regions in the other two patients. In the third patient, fMRI demonstrated an activation in the supplementary motxor area. This study confirms the most significant activation with a high firing rate of the irritative focus, but also suggests the importance of using new techniques (such as EEG-fMRI to examine cerebral blood flow) to identify the controlateral limbic activation, and any other extratemporal activations, possible causes of drug resistance in MTS that may require a more precise pre-surgical evaluation with invasive techniques.

Electrical source imaging of sleep spindles.

To identify and compare cortical source generators of slow and fast sleep spindles in healthy subjects, electroencephalographic (EEG) signals were obtained from 256 channels, and sources on neuroanatomical Montreal Neurological Institute (MNI) space estimated with low-resolution brain electromagnetic tomography analysis (LORETA). Spindle activity was recorded in 18 healthy volunteers during daytime napping. Because of lack of sleep or excessive artifacts, data from 13 subjects were analyzed off-line. Spindles were visually scored, marked, and bandpass filtered (slow 10-12 Hz or fast 12-14 Hz). EEG was segmented on the marker, and segments separately averaged. LORETA projected cortical sources on the MNI brain. Maximal intra- and inter-individual intensities were compared using the Wilcoxon test (P < .05) and cortical sources distribution compared using a chi2 test. Two to three slow spindles generators were consistently identified in frontal lobes, with additional sources in parietal and limbic lobes in half cases. Fast spindles had multiple temporo-parietal sources, with an inconstant frontal source. Inter-individual (P = 0.44), and intra-individual (P = 0.09 slow and P = 0.10 fast spindles) source intensities were comparable. Slow spindles sources were preferentially concentrated over frontal cortices in comparison with fast spindles (P = 0.0009). Our results demonstrate multiple, synchronous, and equipotent spindles cortical generators in healthy subjects, with more anterior generators for slow spindles.

Effect of voluntary repetitive long-lasting muscle contraction activity on the BOLD signal as assessed by optimal hemodynamic response function.

OBJECTIVE: Among other neuroimaging techniques, functional magnetic resonance imaging (fMRI) can be useful for studying the development of motor fatigue. The aim of this study was to identify differences in cortical neuronal activation in nine subjects on three motor tasks: right-hand movement with minimum, maximum, and post-fatigue maximum finger flexion. MATERIALS AND METHODS: fMRI activation maps for each subject and during each condition were obtained by estimating the optimal model of the hemodynamic response function (HRF) out of four standard HRF models and an individual-based HRF model (ibHRF). RESULTS: ibHRF was selected as the optimal model in six out of nine subjects for minimum movement, in five out of nine for maximum movement, and in eight out of nine for post-fatigue maximum movement. As compared to maximum movement, a large reduction in the total number of active voxels (primary sensorimotor area, supplementary motor area and cerebellum) was observed in post-fatigue maximum movement. CONCLUSION: This is the first approach to the evaluation of long-lasting contraction effort in healthy subjects by means of the fMRI paradigm with the use of an individual-based hemodynamic response. The results may be relevant for defining a baseline in future studies on central fatigue in patients with neuropathological disorders.

Reproducibility of EEG-fMRI results in a patient with fixation-off sensitivity.

Blood oxygenation level-dependent (BOLD) activation associated with interictal epileptiform discharges in a patient with fixation-off sensitivity (FOS) was studied using a combined electroencephalography-functional magnetic resonance imaging (EEG-fMRI) technique. An automatic approach for combined EEG-fMRI analysis and a subject-specific hemodynamic response function was used to improve general linear model analysis of the fMRI data. The EEG showed the typical features of FOS, with continuous epileptiform discharges during elimination of central vision by eye opening and closing and fixation; modification of this pattern was clearly visible and recognizable. During all 3 recording sessions EEG-fMRI activations indicated a BOLD signal decrease related to epileptiform activity in the parietal areas. This study can further our understanding of this EEG phenomenon and can provide some insight into the reliability of the EEG-fMRI technique in localizing the irritative zone.

Time-frequency analysis of short-lasting modulation of EEG induced by TMS during wake, sleep deprivation and sleep.

The occurrence of dynamic changes in spontaneous electroencephalogram (EEG) rhythms in the awake state or sleep is highly variable. These rhythms can be externally modulated during transcranial magnetic stimulation (TMS) with a perturbation method to trigger oscillatory brain activity. EEG-TMS co-registration was performed during standard wake, during wake after sleep deprivation and in sleep in six healthy subjects. Dynamic changes in the regional neural oscillatory activity of the cortical areas were characterized using time-frequency analysis based on the wavelet method, and the modulation of induced oscillations were related to different vigilance states. A reciprocal synchronizing/desynchronizing effect on slow and fast oscillatory activity was observed in response to focal TMS after sleep deprivation and sleep. We observed a sleep-related slight desynchronization of alpha mainly over the frontal areas, and a widespread increase in theta synchronization. These findings could be interpreted as proof of the interference external brain stimulation can exert on the cortex, and how this could be modulated by the vigilance state. Potential clinical applications may include evaluation of hyperexcitable states such as epilepsy or disturbed states of consciousness such as minimal consciousness.

Effect of high-frequency repetitive transcranial magnetic stimulation on brain excitability in severely brain-injured patients in minimally conscious or vegetative state.

BACKGROUND: Repetitive transcranial magnetic stimulation (rTMS) induces prolonged functional changes in the cerebral cortex in normal conditions and in altered states of consciousness. Its therapeutic effects have been variously documented. OBJECTIVE: The aim of this study was to investigate the reactivity of electroencephalography (EEG) and the clinical response in six severely brain-injured patients in an altered state of consciousness (minimally conscious state [MCS] or vegetative state [VS]). EEG rhythm and brain excitability were measured before and after a protocol of high-frequency rTMS. METHODS: All six patients underwent clinical and neurophysiological evaluation before rTMS and immediately thereafter. EEG data in resting state were acquired at the beginning of the exam (T0), after rTMS (T1), and 38 min after rTMS (T2). From these data the power values were computed using Fast Fourier Transform. RESULTS: rTMS over the motor cortex induced long-lasting behavioral and neurophysiological modifications in only one patient in MCS. No significant clinical or EEG modifications were detected in any of the other patients, except for changes in motor threshold and motor evoked potential amplitude over the stimulated motor areas. CONCLUSIONS: The main finding of the study is the correlation between EEG reactivity and clinical response after rTMS. Reappearance of fast activity and an increase in slow activity were noted in the one patient with transitory arousal, whereas no significant reliable changes were observed in the other patients showing no clinical reactivity.

Slow spindles' cortical generators overlap with the epileptogenic zone in temporal epileptic patients: an electrical source imaging study.

OBJECTIVE: To determine whether temporal epileptic patients and normal volunteers display similar sleep spindles' cortical generators as determined by electrical source imaging (ESI), and whether such generators overlap in epilepsy patients with the epileptogenic zone identified by ESI. METHODS: Twelve healthy subjects and twelve temporal lobe pharmaco-resistant epileptic patients underwent a 256-channel EEG recording during a daytime nap. Sleep spindles were analyzed off line, distinguishing slow (10-12 Hz) and fast (12-14 Hz) ones, and the final averaged signal was projected onto a MNI (Montreal Neurological Institute) space to localize cortical generators. The same procedure was performed for averaged epileptic spikes, obtaining their cortical source. Intra- and inter-group statistical analyses were conducted. RESULTS: Multiple, concomitant generators were detected in both populations for slow and fast spindles. Slow spindles in epileptics displayed higher source amplitude in comparison to healthy volunteers (Z=0.001), as well as a preferential localization over the affected temporal cortices (p=0.039). Interestingly, at least one of slow spindles' generators overlapped with the epileptogenic zone. CONCLUSION: Slow spindles, but not fast ones, in temporal epilepsy are mainly generated by the affected temporal lobe. SIGNIFICANCE: These results point to the strict relation between sleep and epilepsy and to the possible cognitive implications of spikes arising from memory-encoding brain structures.

Automatic selection of resting-state networks with functional magnetic resonance imaging.

Functional magnetic resonance imaging (fMRI) during a resting-state condition can reveal the co-activation of specific brain regions in distributed networks, called resting-state networks, which are selected by independent component analysis (ICA) of the fMRI data. One of the major difficulties with component analysis is the automatic selection of the ICA features related to brain activity. In this study we describe a method designed to automatically select networks of potential functional relevance, specifically, those regions known to be involved in motor function, visual processing, executive functioning, auditory processing, memory, and the default-mode network. To do this, image analysis was based on probabilistic ICA as implemented in FSL software. After decomposition, the optimal number of components was selected by applying a novel algorithm which takes into account, for each component, Pearson's median coefficient of skewness of the spatial maps generated by FSL, followed by clustering, segmentation, and spectral analysis. To evaluate the performance of the approach, we investigated the resting-state networks in 25 subjects. For each subject, three resting-state scans were obtained with a Siemens Allegra 3 T scanner (NYU data set). Comparison of the visually and the automatically identified neuronal networks showed that the algorithm had high accuracy (first scan: 95%, second scan: 95%, third scan: 93%) and precision (90%, 90%, 84%). The reproducibility of the networks for visual and automatic selection was very close: it was highly consistent in each subject for the default-mode network (≥92%) and the occipital network, which includes the medial visual cortical areas (≥94%), and consistent for the attention network (≥80%), the right and/or left lateralized frontoparietal attention networks, and the temporal-motor network (≥80%). The automatic selection method may be used to detect neural networks and reduce subjectivity in ICA component assessment.

Visual cortex hyperexcitability in idiopathic generalized epilepsies with photosensitivity: a TMS pilot study.

BACKGROUND: The current understanding of the mechanisms underlying photosensitivity is still limited, although most studies point to a hyperexcitability of the visual cortex. METHODS: Using transcranial magnetic stimulation, we determined the resting motor threshold (rMT) and the phosphene threshold (PT) in 33 patients with IGEs (8 with photosensitivity) compared with 12 healthy controls. RESULTS: Eleven controls (92%) reported phosphenes compared with fifteen (46%) patients with idiopathic generalized epilepsy (p=0.015). Phosphenes were reported more frequently among patients with epilepsy with photosensitivity (87.5%) than in patients with active epilepsy without photosensitivity (30.8%) (p=0.038) and patients with epilepsy in remission without photosensitivity (33.3%) (p=0.054); no differences were found between patients with epilepsy with photosensitivity and controls (p=0.648). Resting motor threshold and phosphene threshold were significantly higher among patients with epilepsy (active epilepsy or epilepsy in remission without photosensitivity) compared to healthy controls (p<0.01). Conversely, patients with active epilepsy and photosensitivity had significantly lower values than controls (p=0.03). CONCLUSIONS: The marked decrease in PT and the high phosphene prevalence in patients with IGE with photosensitivity indicate a regional hyperexcitability of the primary visual cortex. Results of this study also suggest that the PT may serve as a biomarker for excitability in patients with IGE and photosensitivity.

Modulation of event-related desynchronization in robot-assisted hand performance: brain oscillatory changes in active, passive and imagined movements.

BACKGROUND: Robot-assisted therapy in patients with neurological disease is an attempt to improve function in a moderate to severe hemiparetic arm. A better understanding of cortical modifications after robot-assisted training could aid in refining rehabilitation therapy protocols for stroke patients. Modifications of cortical activity in healthy subjects were evaluated during voluntary active movement, passive robot-assisted motor movement, and motor imagery tasks performed under unimanual and bimanual protocols. METHODS: Twenty-one channel electroencephalography (EEG) was recorded with a video EEG system in 8 subjects. The subjects performed robot-assisted tasks using the Bi-Manu Track robot-assisted arm trainer. The motor paradigm was executed during one-day experimental sessions under eleven unimanual and bimanual protocols of active, passive and imaged movements. The event-related-synchronization/desynchronization (ERS/ERD) approach to the EEG data was applied to investigate where movement-related decreases in alpha and beta power were localized. RESULTS: Voluntary active unilateral hand movement was observed to significantly activate the contralateral side; however, bilateral activation was noted in all subjects on both the unilateral and bilateral active tasks, as well as desynchronization of alpha and beta brain oscillations during the passive robot-assisted motor tasks. During active-passive movement when the right hand drove the left one, there was predominant activation in the contralateral side. Conversely, when the left hand drove the right one, activation was bilateral, especially in the alpha range. Finally, significant contralateral EEG desynchronization was observed during the unilateral task and bilateral ERD during the bimanual task. CONCLUSIONS: This study suggests new perspectives for the assessment of patients with neurological disease. The findings may be relevant for defining a baseline for future studies investigating the neural correlates of behavioral changes after robot-assisted training in stroke patients.

Frequency and time-frequency analysis of intraoperative ECoG during awake brain stimulation.

Electrocortical stimulation remains the standard for functional brain mapping of eloquent areas to prevent postoperative functional deficits. The aim of this study was to investigate whether the short-train technique (monopolar stimulation) and Penfield's technique (bipolar stimulation) would induce different effects on brain oscillatory activity in awake patients, as quantified by electrocorticography (ECoG). The study population was seven patients undergoing brain tumor surgery. Intraoperative bipolar and monopolar electrical stimulation for cortical mapping was performed during awake surgery. ECoG was recorded using 1 × 8 electrode strip. Spectral estimation was calculated using a parametric approach based on an autoregressive model. Wavelet-based time-frequency analysis was then applied to evaluate the temporal evolution of brain oscillatory activity. Both monopolar and bipolar stimulation produced an increment in delta and a decrease in beta powers for the motor and the sensory channels. These phenomena lasted about 4 s. Comparison between monopolar and bipolar stimulation showed no significant difference in brain activity. Given the importance of quantitative signal analysis for evaluating response accuracy, ECoG recording during electrical stimulation is necessary to characterize the dynamic processes underlying changes in cortical responses in vivo. This study is a preliminary approach to the quantitative analysis of post-stimulation ECoG signals.

Factors predicting functional and cognitive recovery following severe traumatic, anoxic, and cerebrovascular brain damage.

OBJECTIVES: To compare demographic data, clinical data, and rate of functional and cognitive recovery in patients with severe traumatic, cerebrovascular, or anoxic acquired brain injury (ABI) and to identify factors predicting discharge home. PARTICIPANTS: Three hundred twenty-nine patients with severe ABI (192 with traumatic, 104 with cerebrovascular, and 33 with anoxic brain injury). DESIGN: Longitudinal prospective study of inpatients attending the intensive Rehabilitation Department of the "Sacro Cuore" Don Calabria Hospital (Negrar, Verona, Italy). MAIN MEASURES: Etiology, sex, age, rehabilitation admission interval, rehabilitation length of stay, discharge destination, Glasgow Coma Scale, Disability Rating Scale (DRS), Glasgow Outcome Scale, Levels of Cognitive Functioning, and Functional Independence Measure. RESULTS: Predominant etiology was traumatic; male gender was prevalent in all the etiologic groups; patients with traumatic brain injury were younger than the patients in the other groups and had shorter rehabilitation admission interval, greater functional and cognitive outcomes on all considered scales, and a higher frequency of returning home. Patients with anoxic brain injury achieved the lowest grade of functional and cognitive recovery. Age, etiology, and admission DRS score predicted return home. CONCLUSIONS: Patients with traumatic brain injury achieved greater functional and cognitive improvements than patients with cerebrovascular and anoxic ABI. Age, etiology, and admission DRS score can assist in predicting discharge destination.

Tongue biting in epileptic seizures and psychogenic events: an evidence-based perspective.

Tongue biting (TB) may occur both in seizures and in psychogenic non-epileptic events (PNEEs). We undertook a systematic review to determine sensitivity, specificity, and likelihood ratios (LR) of TB. Five studies (222 epilepsy patients and 181 subjects with PNEEs) were included. There was a statistically significant higher prevalence of TB (both without further specifications on site of lesions and lateral TB) in patients with seizures. Pooled accuracy measures of TB (no further specifications) were sensitivity 38%, specificity 75%, pLR 1.479 (95% CI 1.117-1.957), and nLR 0.837 (95% CI 0.736-0.951). Pooled measures of lateral TB were sensitivity 22%, specificity 100%, pLR 21.386 (95% CI 1.325-345.169), and nLR 0.785 (95% CI 0.705-0.875). Only a pooled analysis of data demonstrated a statistically significant pLR for lateral TB. Lateral TB but not 'any' TB has diagnostic significance in distinguishing seizures from PNEEs, supporting the diagnosis of seizures. Tongue biting without further specifications has, therefore, no value in the differential diagnosis between seizures and PNEEs.