Deficiency of metabolic regulator PKM2 activates the pentose phosphate pathway and generates TCF1+ progenitor CD8+ T cells to improve immunotherapy.

TCF1high progenitor CD8+ T cells mediate the efficacy of immunotherapy; however, the mechanisms that govern their generation and maintenance are poorly understood. Here, we show that targeting glycolysis through deletion of pyruvate kinase muscle 2 (PKM2) results in elevated pentose phosphate pathway (PPP) activity, leading to enrichment of a TCF1high progenitor-exhausted-like phenotype and increased responsiveness to PD-1 blockade in vivo. PKM2KO CD8+ T cells showed reduced glycolytic flux, accumulation of glycolytic intermediates and PPP metabolites and increased PPP cycling as determined by 1,2-13C glucose carbon tracing. Small molecule agonism of the PPP without acute glycolytic impairment skewed CD8+ T cells toward a TCF1high population, generated a unique transcriptional landscape and adoptive transfer of agonist-treated CD8+ T cells enhanced tumor control in mice in combination with PD-1 blockade and promoted tumor killing in patient-derived tumor organoids. Our study demonstrates a new metabolic reprogramming that contributes to a progenitor-like T cell state promoting immunotherapy efficacy.

Risk avoidance and social anxiety in adolescence: Examination of event-related potentials and theta-dynamics on the Balloon Risk Avoidance Task.

Adolescents are at relatively high-risk for developing anxiety, particularly social anxiety. A primary hallmark of social anxiety is the impulse to avoid situations that introduce risk. Here, we examined the neural and behavioral correlates of risk avoidance in adolescents (N=59) 11 to 19 years of age. The Balloon Risk Avoidance Task was used with concurrent electroencephalography to measure event-related potentials (frontal P2; late slow-wave; N2, feedback-related negativity, FRN; posterior P3) and oscillatory dynamics (midfrontal theta, 4-7 Hz) in response to unsuccessful and successful risk avoidance conditions. Social anxiety was measured using the Social Phobia and Anxiety Inventory for Children. Results indicated that, across the whole sample, youth exhibited smaller P3, larger FRN, and larger theta responses to unsuccessful risk avoidance. Youth reporting high (compared to low) levels of social anxiety exhibited larger P2, slow-wave, and FRN responses to unsuccessful, compared to successful, risk avoidance. Further, greater social anxiety was associated with reduced theta responses to successful avoidance. Youth with higher levels of social anxiety showed smaller theta responses to both conditions compared to those with low levels of social anxiety. Taken together, the ERP-component differences and weakened theta power in socially anxious youth following unsuccessful avoidance are informative neural correlates for socially anxious youth during risk avoidance.

Computational modeling of social evaluative decision-making elucidates individual differences in adolescent anxiety.

Adolescents experience significant developmental changes during a time of heightened sensitivity to social cues, particularly rejection by peers, which can be especially overwhelming for those with elevated levels of social anxiety. Social evaluative decision-making tasks have been useful in uncovering the neural correlates of information processing biases; however, linking youths' task-based performance to individual differences in psychopathology (e.g., anxiety symptoms) has proven more elusive. Here, we address this weakness with drift diffusion modeling to decompose youths' performance on the social judgment paradigm (SJP) to determine if this approach is useful in discovering individual differences in anxiety symptoms, as well as puberty, age, and sex. A sample of 103 adolescents (55 males, Mage = 14.49, SD = 1.69) completed the SJP and self-report measures of anxiety, as well as self- and parent-reported measures of puberty. The decision threshold parameter, reflecting the amount of evidence needed to make a social evaluative decision, predicted youth self-reported anxiety, above and beyond typical metrics of SJP performance. Our results highlight the potential advantage of parsing task performance according to the underlying cognitive processes. Future research would likely benefit from applying computational modeling approaches to social judgment tasks when attempting to uncover performance-based individual differences in psychopathology.

Behavioral and neural responses to social exclusion in women: the role of facial attractiveness and friendliness.

The behavioral and neural responses to social exclusion were examined in women randomized to four conditions, varying in levels of attractiveness and friendliness. Informed by evolutionary theory, we predicted that being socially excluded by attractive unfriendly women would be more distressing than being excluded by unattractive women, irrespective of their friendliness level. Our results contradicted most of our predictions but provide important insights into women's responses to interpersonal conflict. Accounting for rejection sensitivity, P300 event-related potential amplitudes were largest when women were excluded by unattractive unfriendly women. This may be due to an expectancy violation or an annoyance with being excluded by women low on social desirability. An examination of anger rumination rates by condition suggests the latter. Only attractive women's attractiveness ratings were lowered in the unfriendly condition, indicating they were specifically punished for their exclusionary behavior. Women were more likely to select attractive women to compete against with one exception-they selected the Black attractive opponent less often than the White attractive opponent when presented as unfriendly. Finally, consistent with studies on retaliation in relation to social exclusion, women tended to rate competitors who rejected them as being more rude, more competitive, less attractive, less nice, and less happy than non-competitors. The ubiquity of social exclusion and its pointed emotional and physiological impact on women demands more research on this topic.

Post-event rumination and social anxiety: a systematic review and meta-analysis.

Post-event rumination, the extent to which one engages in persistent, detailed, and negative thinking following social situations, serves as a risk process in the pathophysiology of social anxiety. Although a substantial body of research has assessed post-event rumination and social anxiety, this literature has produced inconsistent results. We conducted a systematic review and meta-analysis to examine whether the magnitude of the association between post-event rumination and social anxiety varied as a function of questionnaire and/or task utilized. We included all studies reporting a correlation between post-event rumination and social anxiety symptomatology. Fisher's z correlation coefficients were calculated through random-effect meta-analyses. Results indicated a moderate association between post-event rumination and social anxiety symptomatology (r = 0.45, p < 0.001, 95%CI [0.40-0.50]). Subgroup meta-analyses indicated that the type of questionnaire used to assess post-event rumination (Q = 44.36, df = 3, p < 0.001) and social anxiety (Q = 26.44, df = 8, p < 0.001), as well as the task conducted prior to assessing post-event rumination (Q = 14.31, df = 2, p < 0.001), influenced the effect size. This study demonstrates a moderate relation between post-event rumination and social anxiety across the anxiety spectrum, illustrating the importance of treatments specifically targeting post-event rumination. Moreover, we highlight the importance of taking care when designing studies to explore relations between post-event rumination and social anxiety.

Screening and Evaluation of Sleep Disturbances and Sleep Disorders in Children and Adolescents.

Sleep disturbances and sleep disorders are prevalent in children/adolescents and have a bidirectional relationship with pediatric medical and mental health disorders. Screening tools and mechanisms for the evaluation and treatment of sleep disturbances and sleep disorders in the pediatric mental health clinic are less well-known; hence, sleep disturbances and disorders are under-recognized in the pediatric clinics. We present specific, validated screening and evaluation tools to identify sleep disturbances and sleep disorders in children/adolescents. We offer guidance related to the use of consumer wearables for sleep assessments and use of sleep telemedicine in pediatric mental health and primary care clinics.

Relationships Between Prenatal Cocaine Exposure, Cannabis-Use Onset and Emotional and Related Characteristics in Young/Emerging Adults.

Background: Exposure to substances in utero may have significant early-life consequences. Less is known about the effects in emerging adulthood, particularly regarding patterns of substance use and related characteristics.Objectives: In this study, we recruited emerging adults, followed since birth, who had been prenatally exposed, or not, to cocaine. Individuals reported on their cannabis, alcohol, and tobacco use, and measures of impulsivity, anhedonia, emotional regulation, and mental health were obtained. Comparisons were made between emerging adults with prenatal cocaine exposure and those without. Correlations were performed between psychological measures and substance use, and regression analyses were conducted to determine potential pathways by which such measures may relate to prenatal exposure or substance use.Results: Individuals with prenatal cocaine exposure (vs. those without) used cannabis at younger ages, reported greater cannabis-use severity, and demonstrated higher impulsivity, state anxiety, and alexithymia. Earlier age of onset of cannabis use was associated with higher impulsivity, state anxiety, alexithymia, and social and physical anhedonia. Cannabis-use age-of-onset mediated the relationship between prenatal cocaine-exposure status and state anxiety and between prenatal cocaine-exposure status and cannabis-use severity in emerging adulthood but not relationships between prenatal cocaine-exposure status and impulsivity or alexithymia in emerging adulthood. Findings suggest that adults with prenatal cocaine exposure may use cannabis at younger ages, which may relate to increased anxiety and more severe use.Conclusions: These findings suggest both mechanisms and possible intervention targets to improve mental health in emerging adults with prenatal cocaine exposure.

Deficiency of metabolic regulator PKM2 activates the pentose phosphate pathway and generates TCF1+ progenitor CD8+ T cells to improve checkpoint blockade.

TCF1high progenitor CD8+ T cells mediate the efficacy of PD-1 blockade, however the mechanisms that govern their generation and maintenance are poorly understood. Here, we show that targeting glycolysis through deletion of pyruvate kinase muscle 2 (PKM2) results in elevated pentose phosphate pathway (PPP) activity, leading to enrichment of a TCF1high central memory-like phenotype and increased responsiveness to PD-1 blockade in vivo. PKM2KO CD8+ T cells showed reduced glycolytic flux, accumulation of glycolytic intermediates and PPP metabolites, and increased PPP cycling as determined by 1,2 13C glucose carbon tracing. Small molecule agonism of the PPP without acute glycolytic impairment skewed CD8+ T cells towards a TCF1high population, generated a unique transcriptional landscape, enhanced tumor control in mice in combination with PD-1 blockade, and promoted tumor killing in patient-derived tumor organoids. Our study demonstrates a new metabolic reprogramming that contributes to a progenitor-like T cell state amenable to checkpoint blockade.

Catch the drift: Depressive symptoms track neural response during more efficient decision-making for negative self-referents.

Background: Adolescence is a time of heightened risk for developing depression and also a critical period for the development and integration of self-identity. Despite this, the relation between the neurophysiological correlates of self-referential processing and major depressive symptoms in youth is not well understood. Here, we leverage computational modeling of the self-referential encoding task (SRET) to identify behavioral moderators of the association between the posterior late positive potential (LPP), an event-related potential associated with emotion regulation, and youth self-reported symptoms of depression. Specifically, within a drift-diffusion framework, we evaluated whether the association between the posterior LPP and youth symptoms of major depression was moderated by drift rate, a parameter reflecting processing efficiency during self-evaluative decisions. Methods: A sample of 106 adolescents, aged 12 to 17 (53% male; Mage = 14.49, SD = 1.70), completed the SRET with concurrent high-density electroencephalography and self-report measures of depression and anxiety. Results: Findings indicated a significant moderation: for youth showing greater processing efficiency (drift rate) when responding to negative compared to positive words, larger posterior LPPs predicted greater depressive symptom severity. Limitations: We relied on a community sample and our study was cross-sectional in nature. Future longitudinal work with clinically depressed youth would be beneficial. Conclusions: Our results suggest a neurobehavioral model of adolescent depression wherein efficient processing of negative information co-occurs with increased demands on affective self-regulation. Our findings also have clinical relevance; youth's neurophysiological response (posterior LPP) and performance during the SRET may serve as a novel target for tracking treatment-related changes in one's self-identity.

Simultaneous quantitative imaging of two PET radiotracers via the detection of positron-electron annihilation and prompt gamma emissions.

In conventional positron emission tomography (PET), only one radiotracer can be imaged at a time, because all PET isotopes produce the same two 511 keV annihilation photons. Here we describe an image reconstruction method for the simultaneous in vivo imaging of two PET tracers and thereby the independent quantification of two molecular signals. This method of multiplexed PET imaging leverages the 350-700 keV range to maximize the capture of 511 keV annihilation photons and prompt γ-ray emission in the same energy window, hence eliminating the need for energy discrimination during reconstruction or for signal separation beforehand. We used multiplexed PET to track, in mice with subcutaneous tumours, the biodistributions of intravenously injected [124I]I-trametinib and 2-deoxy-2-[18F]fluoro-D-glucose, [124I]I-trametinib and its nanoparticle carrier [89Zr]Zr-ferumoxytol, and the prostate-specific membrane antigen (PSMA) and infused PSMA-targeted chimaeric antigen receptor T cells after the systemic administration of [68Ga]Ga-PSMA-11 and [124I]I. Multiplexed PET provides more information depth, gives new uses to prompt γ-ray-emitting isotopes, reduces radiation burden by omitting the need for an additional computed-tomography scan and can be implemented on preclinical and clinical systems without any modifications in hardware or image acquisition software.

Functional connectivity during tic suppression predicts reductions in vocal tics following behavior therapy in children with Tourette syndrome.

BACKGROUND: Comprehensive Behavioral Intervention for Tics (CBIT) is recommended as a first-line treatment for Tourette syndrome in children and adults. While there is strong evidence proving its efficacy, the mechanisms of reduction in tic severity during CBIT are still poorly understood. In a recent study, our group identified a functional brain network involved in tic suppression in children with TS. We reasoned that voluntary tic suppression and CBIT may share some mechanisms and thus we wanted to assess whether functional connectivity during tic suppression was associated with CBIT outcome. METHODS: Thirty-two children with TS, aged 8 to 13 years old, participated in a randomized controlled trial of CBIT v. a treatment-as-usual control condition. EEG was recorded during tic suppression in all participants at baseline and endpoint. We used a source-reconstructed EEG connectivity pipeline to assess functional connectivity during tic suppression. RESULTS: Functional connectivity during tic suppression did not change from baseline to endpoint. However, baseline tic suppression-related functional connectivity specifically predicted the decrease in vocal tic severity from baseline to endpoint in the CBIT group. Supplementary analyses revealed that the functional connectivity between the right superior frontal gyrus and the right angular gyrus was mainly driving this effect. CONCLUSIONS: This study revealed that functional connectivity during tic suppression at baseline predicted reduction in vocal tic severity. These results suggest probable overlap between the mechanisms of voluntary tic suppression and those of behavior therapy for tics.

EEG coherence before and after giving birth.

During pregnancy and the postpartum period, changes in brain volume and in motivational, sensory, cognitive, and emotional processes have been described. However, to date, longitudinal modifications of brain function have been understudied. To explore regional cortical coupling, in pregnancy and at 3 months postpartum, we analyzed resting-state electroencephalographic (EEG) coherence in the delta, theta, alpha1, alpha2, beta1, and beta2 frequency bands across frontal and parietal regions of the maternal brain (Fp1, Fp2, F3, F4, P3, and P4). We found that from pregnancy to the postpartum period, mothers showed less intrahemispheric EEG coherence between the frontal and parietal regions in the alpha1 and alpha2 bands, as well as greater interhemispheric EEG coherence between frontopolar regions in the beta2 band. These changes suggest decreased inhibition of neural circuits. These neurophysiological changes may represent an adaptive process characteristic of motherhood.

The current state-of-the-art in pharmacotherapy for pediatric generalized anxiety disorder.

INTRODUCTION: Anxiety disorders are the most prevalent psychiatric disorders among youth. Among the various anxiety disorders, generalized anxiety disorder is particularly prevalent. Youth with GAD appear at elevated risk of developing other anxiety disorders, mood disorder, and substance use disorders. Functional outcomes of youth with GAD can be improved through early recognition and treatment, thus promoting better longer-term outcomes. AREAS COVERED: The current article summarizes evidence-based state-of-the-art pharmacotherapy for pediatric GAD based on open-label, randomized, and controlled trials. Two electronic databases (PubMed and Scopus) were systematically searched in April 2022 for relevant publications. EXPERT OPINION: The literature supports a combination of psychotherapy and pharmacotherapy as being associated with better outcomes when compared to mono-therapies. While longer-term follow-ups are limited, one such study does challenge this notion. Both selective serotonin reuptake inhibitors (SSRIs) and selective norepinephrine reuptake inhibitors (SNRIs) have been found across studies to have moderate effect sizes in the treatment of pediatric anxiety disorders. SSRIs continue to be a first-line intervention, whereas SNRIs may be considered a second-line treatment. While more evidence is needed, there are emerging data indicating that SSRIs are associated with a more rapid and greater reduction in anxiety symptoms when compared to SNRIs.

NDUFA4L2 reduces mitochondrial respiration resulting in defective lysosomal trafficking in clear cell renal cell carcinoma.

In clear cell renal cell carcinoma (ccRCC), activation of hypoxic signaling induces NADH dehydrogenase (ubiquinone) 1 alpha subcomplex, 4-like 2 (NDUFA4L2) expression. Over 90% of ccRCCs exhibit overexpression of NDUFA4L2, which we previously showed contributes to ccRCC proliferation and survival. The function of NDUFA4L2 in ccRCC has not been fully elucidated. NDUFA4L2 was reported to reduce mitochondrial respiration via mitochondrial complex I inhibition. We found that NDUFA4L2 expression in human ccRCC cells increases the extracellular acidification rate, indicative of elevated glycolysis. Conversely, NDUFA4L2 expression in non-cancerous kidney epithelial cells decreases oxygen consumption rate while increasing extracellular acidification rate, suggesting that a Warburg-like effect is induced by NDUFA4L2 alone. We performed mass-spectrometry (MS)-based proteomics of NDUFA4L2 associated complexes. Comparing RCC4-P (parental) ccRCC cells with RCC4 in which NDUFA4L2 is knocked out by CRISPR-Cas9 (RCC4-KO-643), we identified 3,215 proteins enriched in the NDUFA4L2 immunoprecipitates. Among the top-ranking pathways were "Metabolic Reprogramming in Cancer" and "Glycolysis Activation in Cancer (Warburg Effect)." We also show that NDUFA4L2 enhances mitochondrial fragmentation, interacts with lysosomes, and increases mitochondrial-lysosomal associations, as assessed by high-resolution fluorescence microscopy and live cell imaging. We identified 161 lysosomal proteins, including Niemann-Pick Disease Type C Intracellular Cholesterol Transporters 1 and 2 (NPC1, NPC2), that are associated with NDUFA4L2 in RCC4-P cells. RCC4-P cells have larger and decreased numbers of lysosomes relative to RCC4 NDUFA4L2 knockout cells. These findings suggest that NDUFA4L2 regulates mitochondrial-lysosomal associations and potentially lysosomal size and abundance. Consequently, NDUFA4L2 may regulate not only mitochondrial, but also lysosomal functions in ccRCC.

Modeling brain dynamics and gaze behavior: Starting point bias and drift rate relate to frontal midline theta oscillations.

Frontal midline theta oscillatory dynamics have been implicated as an important neural signature of inhibitory control. However, most proactive cognitive control studies rely on behavioral tasks where individual differences are inferred through button presses. We applied computational modeling to further refine our understanding of theta dynamics in a cued anti-saccade task with gaze-contingent eye tracking. Using a drift diffusion model, increased frontal midline theta power during high-conflict, relative to low-conflict, trials predicted a more conservative style of responding through the starting point (bias). During both high- and low-conflict trials, increases in frontal midline theta also predicted improvements in response efficiency (drift rate). Regression analyses provided support for the importance of the starting point bias, which was associated with frontal midline theta over the course of the task above-and-beyond both drift rate and mean reaction time. Our findings provide a more thorough understanding of proactive gaze control by linking trial-by-trial increases of frontal midline theta to a shift in starting point bias facilitating a more neutral style of responding.

Tumor-intrinsic IRE1α signaling controls protective immunity in lung cancer.

IRE1α-XBP1 signaling is emerging as a central orchestrator of malignant progression and immunosuppression in various cancer types. Employing a computational XBP1s detection method applied to TCGA datasets, we demonstrate that expression of the XBP1s mRNA isoform predicts poor survival in non-small cell lung cancer (NSCLC) patients. Ablation of IRE1α in malignant cells delays tumor progression and extends survival in mouse models of NSCLC. This protective effect is accompanied by alterations in intratumoral immune cell subsets eliciting durable adaptive anti-cancer immunity. Mechanistically, cancer cell-intrinsic IRE1α activation sustains mPGES-1 expression, enabling production of the immunosuppressive lipid mediator prostaglandin E2. Accordingly, restoring mPGES-1 expression in IRE1αKO cancer cells rescues normal tumor progression. We have developed an IRE1α gene signature that predicts immune cell infiltration and overall survival in human NSCLC. Our study unveils an immunoregulatory role for cancer cell-intrinsic IRE1α activation and suggests that targeting this pathway may help enhance anti-tumor immunity in NSCLC.

Frontal EEG asymmetry moderates the relation between borderline personality disorder features and feelings of social rejection in adolescents.

Although associations among borderline personality disorder (BPD), social rejection, and frontal EEG alpha asymmetry scores (FAA, a neural correlate of emotion regulation and approach-withdrawal motivations) have been explored in different studies, relatively little work has examined these relations during adolescence in the same study. We examined whether FAA moderated the relation between BPD features and rejection sensitivity following a validated social exclusion paradigm, Cyberball. A mixed, clinical-community sample of 64 adolescents (females = 62.5%; Mage = 14.45 years; SD = 1.6; range = 11-17 years) completed psychodiagnostic interviews and a self-report measure of BPD (Time 1). Approximately two weeks later (Time 2), participants completed a resting EEG recording followed by Cyberball. FAA moderated the relation between BPD features and overall feelings of rejection following Cyberball: individuals with greater relative left FAA had the highest and lowest feelings of social rejection depending on whether they had high and low BPD feature scores, respectively. Results remained after controlling for age, sex, gender, depression, and BPD diagnosis. These results suggest that FAA may moderate the relation between BPD features and social rejection, and that left frontal brain activity at rest may be differentially associated with those feelings in BPD. Findings are discussed in terms of the link between left frontal brain activity in the regulation and dysregulation of social approach behaviors, characteristic of BPD.

Increased Alpha-Band Connectivity During Tic Suppression in Children With Tourette Syndrome Revealed by Source Electroencephalography Analyses.

BACKGROUND: Tourette syndrome (TS) is a neurodevelopmental disorder involving chronic motor and phonic tics. Most individuals with TS can suppress their tics for at least a short period of time. Yet, the brain correlates of tic suppression are still poorly understood. METHODS: In the current study, high-density electroencephalography was recorded during a resting-state and a tic suppression session in 72 children with TS. Functional connectivity between cortical regions was assessed in the alpha band (8-13 Hz) using an electroencephalography source connectivity method. Graph theory and network-based statistics were used to assess the global network topology and to identify brain regions showing increased connectivity during tic suppression. RESULTS: Graph theoretical analyses revealed distinctive global network topology during tic suppression, relative to rest. Using network-based statistics, we found a subnetwork of increased connectivity during tic suppression (p < .001). That subnetwork encompassed many cortical areas, including the right superior frontal gyrus and the left precuneus, which are involved in the default mode network. We also found a condition-by-age interaction, suggesting age-mediated increases in connectivity during tic suppression. CONCLUSIONS: These results suggest that children with TS suppress their tics through a brain circuit involving distributed cortical regions, many of which are part of the default mode network. Brain connectivity during tic suppression also increases as youths with TS mature. These results highlight a mechanism by which children with TS may control their tics, which could be relevant for future treatment studies.

Computational Modeling of Self-Referential Processing Reveals Domain General Associations with Adolescent Anxiety Symptoms.

What an adolescent thinks about themselves, commonly termed self-referential processing, has significant implications for youth long-term psychological well-being. Self-referential processing plays an important role in anticipatory and reactive processing in social contexts and contributes to symptoms of social anxiety. Previous work examining self-referential processing largely focuses on child and adolescent depression, relying on endorsement and reaction time for positive and negative self-describing adjectives in a self-referential encoding task (SRET). Here, we employ computational methods to interrogate the latent processes underlying choice reaction times to evaluate the fit of several drift-diffusion models of youth SRET performance. A sample of 106 adolescent, aged 12-17 (53% male; Mage = 14.49, SD = 1.70) completed the SRET and self-report measures of anxiety and depression. Our results support the utility of modeling the SRET, where the rate of evidence accumulation (i.e., drift rate) during negative self-referential processing was related to social anxiety above-and-beyond mean task performance. Our regression analyses indicated that youth efficiency in processing of self-referential views was domain general to anxiety, highlighting the importance of assessing both social and physiological anxiety symptoms when predicting SRET performance. The computational modeling results revealed that self-referential views are not uniquely related to depression-related constructs but also facets of anxiety.

Human visual consciousness involves large scale cortical and subcortical networks independent of task report and eye movement activity.

The full neural circuits of conscious perception remain unknown. Using a visual perception task, we directly recorded a subcortical thalamic awareness potential (TAP). We also developed a unique paradigm to classify perceived versus not perceived stimuli using eye measurements to remove confounding signals related to reporting on conscious experiences. Using fMRI, we discovered three major brain networks driving conscious visual perception independent of report: first, increases in signal detection regions in visual, fusiform cortex, and frontal eye fields; and in arousal/salience networks involving midbrain, thalamus, nucleus accumbens, anterior cingulate, and anterior insula; second, increases in frontoparietal attention and executive control networks and in the cerebellum; finally, decreases in the default mode network. These results were largely maintained after excluding eye movement-based fMRI changes. Our findings provide evidence that the neurophysiology of consciousness is complex even without overt report, involving multiple cortical and subcortical networks overlapping in space and time.