Managing electromyogram contamination in scalp recordings: An approach identifying reliable beta and gamma EEG features of psychoses or other disorders.

OBJECTIVE: In publications on the electroencephalographic (EEG) features of psychoses and other disorders, various methods are utilized to diminish electromyogram (EMG) contamination. The extent of residual EMG contamination using these methods has not been recognized. Here, we seek to emphasize the extent of residual EMG contamination of EEG. METHODS: We compared scalp electrical recordings after applying different EMG-pruning methods with recordings of EMG-free data from 6 fully paralyzed healthy subjects. We calculated the ratio of the power of pruned, normal scalp electrical recordings in the six subjects, to the power of unpruned recordings in the same subjects when paralyzed. We produced "contamination graphs" for different pruning methods. RESULTS: EMG contamination exceeds EEG signals progressively more as frequencies exceed 25 Hz and with distance from the vertex. In contrast, Laplacian signals are spared in central scalp areas, even to 100 Hz. CONCLUSION: Given probable EMG contamination of EEG in psychiatric and other studies, few findings on beta- or gamma-frequency power can be relied upon. Based on the effectiveness of current methods of EEG de-contamination, investigators should be able to reanalyze recorded data, reevaluate conclusions from high-frequency EEG data, and be aware of limitations of the methods.

Fast and effective removal of contamination from scalp electrical recordings.

OBJECTIVE: To present a new, automated and fast artefact-removal approach which significantly reduces the effect of contamination in scalp electrical recordings. METHOD: We used spectral and temporal characteristics of different sources recorded during a typical scalp electrical recording in order to improve a fast and effective artefact removal approach. Our experiments show that correlation coefficient and spectral gradient of brain components differ from artefactual components. We trained two binary support vector machine classifiers such that one separates brain components from muscle components, and the other separates brain components from mains power and environmental components. We compared the performance of the proposed approach with seven currently used alternatives on three datasets, measuring mains power artefact reduction, muscle artefact reduction and retention of brain neurophysiological responses. RESULTS: The proposed approach significantly reduces the main power and muscle contamination from scalp electrical recording without affecting brain neurophysiological responses. None of the competitors outperformed the new approach. CONCLUSIONS: The proposed approach is the best choice for artefact reduction of scalp electrical recordings. Further improvements are possible with improved component analysis algorithms. SIGNIFICANCE: This paper provides a definitive answer to an important question: Which artefact removal algorithm should be used on scalp electrical recordings?

Resting cranial and upper cervical muscle activity is increased in patients with migraine.

OBJECTIVE: To compare comprehensive measures of scalp-recorded muscle activity in migraineurs and controls. METHOD: We used whole-of-head high-density scalp electrical recordings, independent component analysis (ICA) and spectral slope of the derived components, to define muscle (electromyogram-containing) components. After projecting muscle components back to scalp, we quantified scalp spectral power in the frequency range, 52-98 Hz, reflecting muscle activation. We compared healthy subjects (n = 65) and migraineurs during a non-headache period (n = 26). We also examined effects due to migraine severity, gender, scalp-region and task (eyes-closed and eyes-open). We could not examine the effect of pre-ictal versus inter-ictal versus post-ictal as this information was not available in the pre-existing dataset. RESULTS: There was more power due to muscle activity (mean ±â€¯SEM) in migraineurs than controls (respectively, -13.61 ±â€¯0.44 dB versus -14.73 ±â€¯0.24 dB, p = 0.028). Linear regression showed no relationship between headache frequency and muscle activity in any combination of region and task. There was more power during eyes-open than eyes-closed (respectively, -13.42 ±â€¯0.34 dB versus -14.92 ±â€¯0.34 dB, p = 0.002). CONCLUSIONS: There is an increase in cranial and upper cervical muscle activity in non-ictal migraineurs versus controls. This raises questions of the role of muscle in migraine, and the possible differentiation of non-ictal phases. SIGNIFICANCE: This provides preliminary evidence to date of possible cranial muscle involvement in migraine.

Improved artefact removal from EEG using Canonical Correlation Analysis and spectral slope.

BACKGROUND: Contamination of scalp measurement by tonic muscle artefacts, even in resting positions, is an unavoidable issue in EEG recording. These artefacts add significant energy to the recorded signals, particularly at high frequencies. To enable reliable interpretation of subcortical brain activity, it is necessary to detect and discard this contamination. NEW METHOD: We introduce a new automatic muscle-removal approach based on the traditional Blind Source Separation-Canonical Correlation Analysis (BSS-CCA) method and the spectral slope of its components. We show that CCA-based muscle-removal methods can discriminate between signals with high correlation coefficients (brain, mains artefact) and signals with low correlation coefficients (white noise, muscle). We also show that typical BSS-CCA components are not purely from one source, but are mixtures from multiple sources, limiting the performance of BSS-CCA in artefact removal. We demonstrate, using our paralysis dataset, improved performance using BSS-CCA followed by spectral-slope rejection. RESULT: This muscle removal approach can reduce high-frequency muscle contamination of EEG, especially at peripheral channels, while preserving steady-state brain responses in cognitive tasks. COMPARISON WITH EXISTING METHODS: This approach is automatic and can be applied on any sample of data easily. The results show its performance is comparable with the ICA method in removing muscle contamination and has significantly lower computational complexity. CONCLUSION: We identify limitations of the traditional BSS-CCA approach to artefact removal in EEG, propose and test an extension based on spectral slope that makes it automatic and improves its performance, and results in performance comparable to competitors such as ICA-based artefact removal.

Evaluation of a minimum-norm based beamforming technique, sLORETA, for reducing tonic muscle contamination of EEG at sensor level.

BACKGROUND: Cranial and cervical muscle activity (electromyogram, EMG) contaminates the surface electroencephalogram (EEG) from frequencies below 20 through to frequencies above 100Hz. It is not possible to have a reliable measure of cognitive tasks expressed in EEG at gamma-band frequencies until the muscle contamination is removed. NEW METHOD: In the present work, we introduce a new approach of using a minimum-norm based beamforming technique (sLORETA) to reduce tonic muscle contamination at sensor level. Using a generic volume conduction model of the head, which includes three layers (brain, skull, and scalp), and sLORETA, we estimated time-series of sources distributed within the brain and scalp. The sources within the scalp were considered to be muscle and discarded in forward modelling. RESULT: (1) The method reduced EMG contamination, more strongly at peripheral channels; (2) task-induced cortical activity was retained or revealed after removing putative muscle activity. COMPARISON WITH EXISTING METHODS: This approach can decrease tonic muscle contamination in scalp measurements without relying on time-consuming processing of expensive MRI data. In addition, it is competitive to ICA in muscle reduction and can be reliably applied on any length of recorded data that captures the dynamics of the signals of interest. CONCLUSION: This study suggests that sLORETA can be used as a method to quantitate cranial muscle activity and reduce its contamination at sensor level.

Partial duplication of the CRYBB1-CRYBA4 locus is associated with autosomal dominant congenital cataract.

Congenital cataract is a rare but severe paediatric visual impediment, often caused by variants in one of several crystallin genes that produce the bulk of structural proteins in the lens. Here we describe a pedigree with autosomal dominant isolated congenital cataract and linkage to the crystallin gene cluster on chromosome 22. No rare single nucleotide variants or short indels were identified by exome sequencing, yet copy number variant analysis revealed a duplication spanning both CRYBB1 and CRYBA4. While the CRYBA4 duplication was complete, the CRYBB1 duplication was not, with the duplicated CRYBB1 product predicted to create a gain of function allele. This association suggests a new genetic mechanism for the development of isolated congenital cataract.

Electroencephalographic correlates of states of concentrative meditation.

Meditative techniques aim for and meditators report states of mental alertness and focus, concurrent with physical and emotional calm. We aimed to determine the electroencephalographic (EEG) correlates of five states of Buddhist concentrative meditation, particularly addressing a correlation with meditative level. We studied 12 meditators and 12 pair-matched meditation-naïve participants using high-resolution scalp-recorded EEG. To maximise reduction of EMG, data were pre-processed using independent component analysis and surface Laplacian transformed data. Two non-meditative and five meditative states were used: resting baseline, mind-wandering, absorptions 1, 2, 3, 4 and 5 (corresponding to four levels of absorption and an absorption with a different object of focus, otherwise equivalent to level 4; these five meditative states produce repeatable, distinctly different experiences for experienced meditators). The experimental protocol required participants to experience the states in the order listed above, followed immediately by the reverse. We then calculated EEG power in standard frequency bands from 1 to 80Hz. We observed decreases of central scalp beta (13-25Hz), and central low gamma (25-48Hz) power in meditators during deeper absorptions. In contrast, we identified increases in frontal midline and temporo-parietal theta power in meditators, again, during deeper absorptions. Alpha activity was increased over all meditative states, not depth-related. This study demonstrates that the subjective experiences of deepening meditation partially correspond to measures of EEG. Our results are in accord with prior studies on non-graded meditative states. These results are also consistent with increased theta correlating with tightness of focus, and reduced beta/gamma with the desynchronization associated with enhanced alertness.

A bundled care approach to patients with idiopathic pulmonary fibrosis improves transplant-free survival.

BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a chronic lung disease with poor prognosis and limited therapeutic options. The 2011 ATS/ERS/JRS/ALAT consensus statement provided a number of recommendations for the management of IPF patients. The primary objective of this study was to determine if "bundling" these recommendations in the management of patients with IPF impacts clinical outcomes. METHODS: We conducted a single center, retrospective cohort study of 284 patients diagnosed with IPF. The proposed bundle of care (BOC) components were: (1) visits to a specialized interstitial lung diseases clinic with evaluation of pulmonary function tests at least twice yearly; (2) referral to pulmonary rehabilitation yearly; (3) timed walk test yearly; (4) echocardiogram yearly; and (5) gastroesophageal reflux therapy. Each component of the BOC was given a score of "1" per year of follow up, and the average sum of the scores (ranging from 0 to 5) was determined for the entire period of follow-up (BOCS), as well as during the first year of follow-up (BOCY1). The primary outcome measure was transplant-free survival. RESULTS: Age, gender, smoking status, BMI, %FVC, %DLCO did not differ between levels of BOCS and BOCY1. Lowest BOCS (≤1) was associated with a lower transplant-free survival independent of age and %FVC compared to patients with the highest BOCS (>4) (HR 2.274, CI 1.12-4.64, p = 0.024). Lower BOCY1 was associated with a higher risk for transplant or death independent of age and %FVC in comparison to patients with highest BOCY1 (≤1 vs. >4, HR 2.23, p = 0.014; >1 to 2 vs. >4, HR 1.87, p = 0.011; >2 to 3 vs. >4, HR 1.72, p = 0.019). CONCLUSION: IPF patients with higher BOC scores had improved transplant-free survival. Prospective studies are needed to confirm these findings and determine the best strategies for the management of patients with IPF.

A comparison of independent component analysis algorithms and measures to discriminate between EEG and artifact components.

Independent Component Analysis (ICA) is a powerful statistical tool capable of separating multivariate scalp electrical signals into their additive independent or source components, specifically EEG or electroencephalogram and artifacts. Although ICA is a widely accepted EEG signal processing technique, classification of the recovered independent components (ICs) is still flawed, as current practice still requires subjective human decisions. Here we build on the results from Fitzgibbon et al. [1] to compare three measures and three ICA algorithms. Using EEG data acquired during neuromuscular paralysis, we tested the ability of the measures (spectral slope, peripherality and spatial smoothness) and algorithms (FastICA, Infomax and JADE) to identify components containing EMG. Spatial smoothness showed differentiation between paralysis and pre-paralysis ICs comparable to spectral slope, whereas peripherality showed less differentiation. A combination of the measures showed better differentiation than any measure alone. Furthermore, FastICA provided the best discrimination between muscle-free and muscle-contaminated recordings in the shortest time, suggesting it may be the most suited to EEG applications of the considered algorithms. Spatial smoothness results suggest that a significant number of ICs are mixed, i.e. contain signals from more than one biological source, and so the development of an ICA algorithm that is optimised to produce ICs that are easily classifiable is warranted.

Constitutive spectral EEG peaks in the gamma range: suppressed by sleep, reduced by mental activity and resistant to sensory stimulation.

OBJECTIVE: In a systematic study of gamma activity in neuro-psychiatric disease, we unexpectedly observed distinctive, apparently persistent, electroencephalogram (EEG) spectral peaks in the gamma range (25-100 Hz). Our objective, therefore, was to examine the incidence, distribution and some of the characteristics of these peaks. METHODS: High sample-rate, 128-channel, EEG was recorded in 603 volunteers (510 with neuropsychiatric disorders, 93 controls), whilst performing cognitive tasks, and converted to power spectra. Peaks of spectral power, including in the gamma range, were determined algorithmically for all electrodes. To determine if peaks were stable, 24-h ambulatory recordings were obtained from 16 subjects with peaks. In 10 subjects, steady-state responses to stimuli at peak frequency were compared with off-peak-frequency stimulation to determine if peaks were a feature of underlying network resonances and peaks were evaluated with easy and hard versions of oddball tasks to determine if peaks might be influenced by mental effort. RESULTS: 57% of 603 subjects exhibited peaks >2 dB above trough power at or above 25 Hz. Larger peaks (>5 dB) were present in 13% of subjects. Peaks were distributed widely over the scalp, more frequent centrally. Peaks were present through the day and were suppressed by slow-wave-sleep. Steady-state responses were the same with on- or off-peak sensory stimulation. In contrast, mental effort resulted in reductions in power and frequency of gamma peaks, although the suppression did not correlate with level of effort. CONCLUSIONS: Gamma EEG can be expressed constitutively as concentrations of power in narrow or wide frequency bands that play an, as yet, unknown role in cognitive activity. SIGNIFICANCE: These findings expand the described range of rhythmic EEG phenomena. In particular, in addition to evoked, induced and sustained gamma band activity, gamma activity can be present constitutively in spectral peaks.

Spreading depression: evidence of five electroencephalogram phases.

Spreading depression (SD), a self-propagating wave of astroglial and neuronal depolarization, is an accompaniment of several neurological disorders including epilepsy. Its well-described features are initial depolarization, followed by EEG flattening. In this in vivo study in awake animals, the relationship of SDs to epileptiform activity was re-examined. We assessed SDs generated by mechanical stimulation and by metabolic inhibition with fluorocitrate. In addition to identifying prolonged EEG depression, we identified two periods, one prior to and another during depression, characterized by increases in power of specific frequencies that were sometimes associated with epileptiform discharges. The first period was characterized by ripple activity close to the induction site (88% of SDs with intracortical electrodes). The second period was characterized by localized low-frequency spikes (100% with dural screw electrodes, 65% with intracortical electrodes). By using fluorocitrate to induce SDs, the initial period was also characterized by runs of spikes (52%). Finally, with SDs induced by both methods, there was a period at the end of depression when additional, unprovoked SDs occurred (20%). Five stages of SD were defined by these phenomena, in the order: excitation, depression, excitation, depression, SD, with metabolic inhibition enhancing the expression of epileptiform spiking.

Two types of transgenic lines for doxycycline-inducible, cell-specific gene expression in zebrafish ultraviolet cone photoreceptors.

Temporal and spatial control of gene expression is important for studying the molecular and cellular mechanisms of development, physiology, and disease. We used the doxycycline (Dox)-inducible, Tet-On system to develop transgenic zebrafish for inducible, cell specific control of gene expression in the ultraviolet (UV) cone photoreceptors. Two constructs containing the reverse tetracycline-controlled transcriptional transactivator (rtTA) gene driven by the UV opsin-specific promoter (opn1sw1) were used to generate stable transgenic zebrafish lines using the Tol2-based transgenesis method. One construct included a self-reporting GFP (opn1sw1:rtTA, TRE:GFP) and the other incorporated an epitope tag on the rtTA protein (opn1sw1:rtTA(flag)). UV cone-specific expression of TRE-controlled transgenes was induced by Dox treatment in larvae and adults. Induction of gene expression was observed in 96% of all larval UV cones within 16 h of Dox treatment. UV cone-specific expression of two genes from a bidirectional TRE construct injected into one-cell Tg(opn1sw1:rtTA(flag)) embryos were also induced by Dox treatment. In addition, UV cone-specific expression of Crb2a(IntraWT) was induced by Dox treatment in progeny from crosses of the TRE-response transgenic line, Tg(TRE:HA-Crb2a(IntraWT)), to the Tg(opn1sw1:rtTA, TRE:GFP) line and the Tg(opn1sw1:rtTA(flag)) line. These lines can be used in addition to the inducible, rod-specific gene expression system from the Tet-On Toolkit to elucidate the photoreceptor-specific effects of genes of interest in photoreceptor cell biology and retinal disease.

Surface Laplacian of central scalp electrical signals is insensitive to muscle contamination.

The objective of this paper was to investigate the effects of surface Laplacian processing on gross and persistent electromyographic (EMG) contamination of electroencephalographic (EEG) signals in electrical scalp recordings. We made scalp recordings during passive and active tasks, on awake subjects in the absence and in the presence of complete neuromuscular blockade. Three scalp surface Laplacian estimators were compared to left ear and common average reference (CAR). Contamination was quantified by comparing power after paralysis (brain signal, B) with power before paralysis (brain plus muscle signal, B+M). Brain:Muscle (B:M) ratios for the methods were calculated using B and differences in power after paralysis to represent muscle (M). There were very small power differences after paralysis up to 600 Hz using surface Laplacian transforms (B:M > 6 above 30 Hz in central scalp leads). Scalp surface Laplacian transforms reduce muscle power in central and pericentral leads to less than one sixth of the brain signal, two to three times better signal detection than CAR. Scalp surface Laplacian transformations provide robust estimates for detecting high-frequency (gamma) activity, for assessing electrophysiological correlates of disease, and also for providing a measure of brain electrical activity for use as a standard in the development of brain/muscle signal separation methods.

Generation of a genetically encoded marker of rod photoreceptor outer segment growth and renewal.

Vertebrate photoreceptors are specialized light sensing neurons. The photoreceptor outer segment is a highly modified cilium where photons of light are transduced into a chemical and electrical signal. The outer segment has the typical cilary axoneme but, in addition, it has a large number of densely packed, stacked, intramembranous discs. The molecular and cellular mechanisms that contribute to vertebrate photoreceptor outer segment morphogenesis are still largely unknown. Unlike typical cilia, the outer segment is continuously regenerated or renewed throughout the life of the animal through the combined process of distal outer segment shedding and proximal outer segment growth. The process of outer segment renewal was discovered over forty years ago, but we still lack an understanding of how photoreceptors renew their outer segments and few, if any, molecular mechanisms that regulate outer segment growth or shedding have been described. Our lack of progress in understanding how photoreceptors renew their outer segments has been hampered by the difficulty in measuring rates of renewal. We have created a new method that uses heat-shock induction of a fluorescent protein that can be used to rapidly measure outer segment growth rates. We describe this method, the stable transgenic line we created, and the growth rates observed in larval and adult rod photoreceptors using this new method. This new method will allow us to begin to define the genetic and molecular mechanisms that regulate rod outer segment renewal, a crucial aspect of photoreceptor function and, possibly, viability.

Two types of Tet-On transgenic lines for doxycycline-inducible gene expression in zebrafish rod photoreceptors and a gateway-based tet-on toolkit.

The ability to control transgene expression within specific tissues is an important tool for studying the molecular and cellular mechanisms of development, physiology, and disease. We developed a Tet-On system for spatial and temporal control of transgene expression in zebrafish rod photoreceptors. We generated two transgenic lines using the Xenopus rhodopsin promoter to drive the reverse tetracycline-controlled transcriptional transactivator (rtTA), one with self-reporting GFP activity and one with an epitope tagged rtTA. The self-reporting line includes a tetracycline response element (TRE)-driven GFP and, in the presence of doxycycline, expresses GFP in larval and adult rods. A time-course of doxycycline treatment demonstrates that maximal induction of GFP expression, as determined by the number of GFP-positive rods, is reached within approximately 24 hours of drug treatment. The epitope-tagged transgenic line eliminates the need for the self-reporting GFP activity by expressing a FLAG-tagged rtTA protein. Both lines demonstrate strong induction of TRE-driven transgenes from plasmids microinjected into one-cell embryos. These results show that spatial and temporal control of transgene expression can be achieved in rod photoreceptors. Additionally, system components are constructed in Gateway compatible vectors for the rapid cloning of doxycycline-inducible transgenes and use in other areas of zebrafish research.

Visual Experiences during Paralysis.

RATIONALE: Paralyzed human volunteers (n = 6) participated in several studies the primary one of which required full neuromuscular paralysis while awake. After the primary experiment, while still paralyzed and awake, subjects undertook studies of humor and of attempted eye-movement. The attempted eye-movements tested a central, intentional component to one's internal visual model and are the subject of this report. METHODS: Subjects reclined in a supportive chair and were ventilated after paralysis (cisatracurium, 20 mg intravenously). In illumination, subjects were requested to focus alternately on the faces of investigators standing on the left and the right within peripheral vision. In darkness, subjects were instructed to look away from a point source of light. Subjects were to report their experiences after reversal of paralysis. RESULTS: During attempted eye-movement in illumination, one subject had an illusion of environmental movement but four subjects perceived faces as clearly as if they were in central vision. In darkness, four subjects reported movement of the target light in the direction of attempted eye-movements and three could control the movement of the light at will. CONCLUSION: The hypothesis that internal visual models receive intended ocular-movement-information directly from oculomotor centers is strengthened by this evidence.

EEG spindles in the rat: evidence for a synchronous network phenomenon.

Whether physiological and pathological electroencephalographic (EEG) spindling phenomena are generated by similar mechanisms remains unknown. We studied four different types of spindling events analysing the behaviour of 19 brain regions during these events in the intact brain of freely behaving rats hypothesising similar generation with a leading role of thalamus or cortex for the dominant rhythm of the spindles with a hierarchy of time delays for propagation throughout the brain led by these structures. Autoregressive analysis revealed that all structures studied were capable of leading a spindle, although some did so infrequently. The highest incidence of significant leadership was shown by hindlimb cortex with a proportion of 0.38 during picrotoxin spindles. Contralateral time delays within bilateral structures were extremely small demonstrating synchronous activity with no dominant hemisphere for any of the spindling activities (range: 0.1+/-0.4 ms for frontal cortex to 10.0+/-1.6 ms for piriform cortex). Between different structures, time delays demonstrated that spindling activity occurred in all structures almost simultaneously (range: 0.2+/-1.4 ms between frontal cortex and piriform cortex to 12.3+/-1.2 ms between hindlimb cortex and amygdala). We conclude that different spindle types appear to be subtle alterations of a single phenomenon, during which many brain regions briefly synchronise.

Relation of gamma oscillations in scalp recordings to muscular activity.

We recorded scalp electrical activity before and after full neuro-muscular paralysis in 5 volunteers and determined differences due to elimination of muscular activity on several standard applications of EEG. Due to paralysis, there were reductions in 'noisiness' of the standard scalp recordings which were maximal over the peripheral scalp, not explained by abolition of movement artefact, and best accounted for by sustained EMG activity in resting individuals. There was a corresponding reduction in spectral power in the gamma range. In central leads, the extent of gamma frequency coherence during a non-time-locked mental task (1 s epochs) was reduced by paralysis, likely due to a reduction in gamma-frequency coherence in widely arising EMG signals. In a time-locked mental task (auditory oddball), evoked responses were qualitatively unaffected by paralysis but 3 of 4 induced gamma responses were obscured by EMG.