Feasibility, efficacy, and functional relevance of automated auditory closed-loop suppression of slow-wave sleep in humans.

Slow-wave sleep (SWS) is a fundamental physiological process, and its modulation is of interest for basic science and clinical applications. However, automatised protocols for the suppression of SWS are lacking. We describe the development of a novel protocol for the automated detection (based on the whole head topography of frontal slow waves) and suppression of SWS (through closed-loop modulated randomised pulsed noise), and assessed the feasibility, efficacy and functional relevance compared to sham stimulation in 15 healthy young adults in a repeated-measure sleep laboratory study. Auditory compared to sham stimulation resulted in a highly significant reduction of SWS by 30% without affecting total sleep time. The reduction of SWS was associated with an increase in lighter non-rapid eye movement sleep and a shift of slow-wave activity towards the end of the night, indicative of a homeostatic response and functional relevance. Still, cumulative slow-wave activity across the night was significantly reduced by 23%. Undisturbed sleep led to an evening to morning reduction of wake electroencephalographic theta activity, thought to reflect synaptic downscaling during SWS, while suppression of SWS inhibited this dissipation. We provide evidence for the feasibility, efficacy, and functional relevance of a novel fully automated protocol for SWS suppression based on auditory closed-loop stimulation. Future work is needed to further test for functional relevance and potential clinical applications.

Closed-loop modulation of local slow oscillations in human NREM sleep.

Slow-wave sleep is the deep non-rapid eye-movement (NREM) sleep stage that is most relevant for the recuperative function of sleep. Its defining property is the presence of slow oscillations (<2 Hz) in the scalp electroencephalogram (EEG). Slow oscillations are generated by a synchronous back and forth between highly active UP-states and silent DOWN-states in neocortical neurons. Growing evidence suggests that closed-loop sensory stimulation targeted at UP-states of EEG-defined slow oscillations can enhance the slow oscillatory activity, increase sleep depth, and boost sleep's recuperative functions. However, several studies failed to replicate such findings. Failed replications might be due to the use of conventional closed-loop stimulation algorithms that analyze the signal from one single electrode and thereby neglect the fact that slow oscillations vary with respect to their origins, distributions, and trajectories on the scalp. In particular, conventional algorithms nonspecifically target functionally heterogeneous UP-states of distinct origins. After all, slow oscillations at distinct sites of the scalp have been associated with distinct functions. Here we present a novel EEG-based closed-loop stimulation algorithm that allows targeting UP- and DOWN-states of distinct cerebral origins based on topographic analyses of the EEG: the topographic targeting of slow oscillations (TOPOSO) algorithm. We present evidence that the TOPOSO algorithm can detect and target local slow oscillations with specific, predefined voltage maps on the scalp in real-time. When compared to a more conventional, single-channel-based approach, TOPOSO leads to fewer but locally more specific stimulations in a simulation study. In a validation study with napping participants, TOPOSO targets auditory stimulation reliably at local UP-states over frontal, sensorimotor, and centro-parietal regions. Importantly, auditory stimulation temporarily enhanced the targeted local state. However, stimulation then elicited a standard frontal slow oscillation rather than local slow oscillations. The TOPOSO algorithm is suitable for the modulation and the study of the functions of local slow oscillations.

Sleep-learning impairs subsequent awake-learning.

Although we can learn new information while asleep, we usually cannot consciously remember the sleep-formed memories - presumably because learning occurred in an unconscious state. Here, we ask whether sleep-learning expedites the subsequent awake-learning of the same information. To answer this question, we reanalyzed data (Züst et al., 2019, Curr Biol) from napping participants, who learned new semantic associations between pseudowords and translation-words (guga-ship) while in slow-wave sleep. They retrieved sleep-formed associations unconsciously on an implicit memory test following awakening. Then, participants took five runs of paired-associative learning to probe carry-over effects of sleep-learning on awake-learning. Surprisingly, sleep-learning diminished awake-learning when participants learned semantic associations that were congruent to sleep-learned associations (guga-boat). Yet, learning associations that conflicted with sleep-learned associations (guga-coin) was unimpaired relative to learning new associations (resun-table; baseline). We speculate that the impeded wake-learning originated in a deficient synaptic downscaling and resulting synaptic saturation in neurons that were activated during both sleep-learning and awake-learning.

Skepticism toward Emerging Infectious Diseases and Influenza Vaccination Intentions in Nurses.

Nurses generally show low compliance with vaccination recommendations. We assessed whether low vaccine acceptance is due to skeptical attitudes toward emerging infectious diseases (EIDs). Skepticism toward EIDs manifests as doubts about the real threat of emerging diseases and as distrust in the motives and the competence of institutions that fight these diseases. We performed a cross-sectional questionnaire study in 293 Swiss nurses using a newly developed scale to assess skepticism toward EIDs. Skepticism affected nurses' intentions to vaccinate themselves against seasonal influenza and against possible future pandemic influenza. The influence of skepticism persisted after controlling for other factors that are known to determine nurses' vaccination behavior, namely vaccination habits, feeling at risk of catching influenza, and perceiving vaccination as a professional duty. Skeptical attitudes toward EIDs seem to have a unique and hitherto ignored impact on vaccination intentions. Nurses' vaccine acceptance could be increased if vaccination campaigns specifically target skeptical attitudes toward EIDs. These campaigns should address nurses' doubts about the real threat of EIDs and should rebuild their trust in institutions which fight these diseases.

Unconscious processing effects manifest only if conscious processing is excluded.

In their discussion paper Steinkrauss and Slotnick argue against a role for the hippocampus in unconscious memory formation and retrieval. Unfortunately, they omitted highly relevant evidence that supports a role for the hippocampus in unconscious memory. They criticize four articles, two from our laboratory, pointing out long-known confounds like residual consciousness. We uncover these reproaches as untrue allegations. In our own interest, we prevented conscious mnemonic processing because reliable unconscious memory effects manifest only if consciousness is completely excluded, and because we always knew that residual consciousness would be our Achilles heel for the proponents of the 'explicit memory dogma.'

Episodic long-term memory formation during slow-wave sleep.

We are unresponsive during slow-wave sleep but continue monitoring external events for survival. Our brain wakens us when danger is imminent. If events are non-threatening, our brain might store them for later consideration to improve decision-making. To test this hypothesis, we examined whether novel vocabulary consisting of simultaneously played pseudowords and translation words are encoded/stored during sleep, and which neural-electrical events facilitate encoding/storage. An algorithm for brain-state-dependent stimulation selectively targeted word pairs to slow-wave peaks or troughs. Retrieval tests were given 12 and 36 hr later. These tests required decisions regarding the semantic category of previously sleep-played pseudowords. The sleep-played vocabulary influenced awake decision-making 36 hr later, if targeted to troughs. The words' linguistic processing raised neural complexity. The words' semantic-associative encoding was supported by increased theta power during the ensuing peak. Fast-spindle power ramped up during a second peak likely aiding consolidation. Hence, new vocabulary played during slow-wave sleep was stored and influenced decision-making days later.

Inverse forgetting in unconscious episodic memory.

Forming memories of experienced episodes calls upon the episodic memory system. Episodic encoding may proceed with and without awareness of episodes. While up to 60% of consciously encoded episodes are forgotten after 10 h, the fate of unconsciously encoded episodes is unknown. Here we track over 10 h, which are filled with sleep or daytime activities, the retention of unconsciously and consciously experienced episodes. The episodes were displayed in cartoon clips that were presented weakly and strongly masked for conscious and unconscious encoding, respectively. Clip retention was tested for distinct clips directly after encoding, 3 min and 10 h after encoding using a forced-choice test that demands deliberate responses in both consciousness conditions. When encoding was conscious, retrieval accuracy decreased by 25% from 3 min to 10 h, irrespective of sleep or wakefulness. When encoding was unconscious, retrieval accuracy increased from 3 min to 10 h and depended on sleep. Hence, opposite to the classic forgetting curve, unconsciously acquired episodic memories strengthen over time and hinge on sleep on the day of learning to gain influence over human behavior.

Bi-Temporal Anodal Transcranial Direct Current Stimulation during Slow-Wave Sleep Boosts Slow-Wave Density but Not Memory Consolidation.

Slow-wave sleep (SWS) has been shown to promote long-term consolidation of episodic memories in hippocampo-neocortical networks. Previous research has aimed to modulate cortical sleep slow-waves and spindles to facilitate episodic memory consolidation. Here, we instead aimed to modulate hippocampal activity during slow-wave sleep using transcranial direct current stimulation in 18 healthy humans. A pair-associate episodic memory task was used to evaluate sleep-dependent memory consolidation with face-occupation stimuli. Pre- and post-nap retrieval was assessed as a measure of memory performance. Anodal stimulation with 2 mA was applied bilaterally over the lateral temporal cortex, motivated by its particularly extensive connections to the hippocampus. The participants slept in a magnetic resonance (MR)-simulator during the recordings to test the feasibility for a future MR-study. We used a sham-controlled, double-blind, counterbalanced randomized, within-subject crossover design. We show that stimulation vs. sham significantly increased slow-wave density and the temporal coupling of fast spindles and slow-waves. While retention of episodic memories across sleep was not affected across the entire sample of participants, it was impaired in participants with below-average pre-sleep memory performance. Hence, bi-temporal anodal direct current stimulation applied during sleep enhanced sleep parameters that are typically involved in memory consolidation, but it failed to improve memory consolidation and even tended to impair consolidation in poor learners. These findings suggest that artificially enhancing memory-related sleep parameters to improve memory consolidation can actually backfire in those participants who are in most need of memory improvement.

Larger capacity for unconscious versus conscious episodic memory.

Episodic memory is the memory for experienced events. A peak competence of episodic memory is the mental combination of events to infer commonalities. Inferring commonalities may proceed with and without consciousness of events. Yet what distinguishes conscious from unconscious inference? This question inspired nine experiments that featured strongly and weakly masked cartoon clips presented for unconscious and conscious inference. Each clip featured a scene with a visually impenetrable hiding place. Five animals crossed the scene one-by-one consecutively. One animal trajectory represented one event. The animals moved through the hiding place, where they might linger or not. The participants' task was to observe the animals' entrances and exits to maintain a mental record of which animals hid simultaneously. We manipulated information load to explore capacity limits. Memory of inferences was tested immediately, 3.5 or 6 min following encoding. The participants retrieved inferences well when encoding was conscious. When encoding was unconscious, the participants needed to respond intuitively. Only habitually intuitive decision makers exhibited a significant delayed retrieval of inferences drawn unconsciously. Their unconscious retrieval performance did not drop significantly with increasing information load, while conscious retrieval performance dropped significantly. A working memory network, including hippocampus, was activated during both conscious and unconscious inference and correlated with retrieval success. An episodic retrieval network, including hippocampus, was activated during both conscious and unconscious retrieval of inferences and correlated with retrieval success. Only conscious encoding/retrieval recruited additional brain regions outside these networks. Hence, levels of consciousness influenced the memories' behavioral impact, memory capacity, and the neural representational code.

Learning During Sleep: A Dream Comes True?

Can information that is processed during sleep influence awake behavior? Recent research demonstrates that learning during sleep is possible, but that sleep-learning invariably produces memory traces that are consciously inaccessible in the awake state. Thus, sleep-learning can likely exert implicit, but not explicit, influences on awake behavior.

Implicit Vocabulary Learning during Sleep Is Bound to Slow-Wave Peaks.

Learning while asleep is a dream of mankind, but is often deemed impossible because sleep lacks the conscious awareness and neurochemical milieu thought to be necessary for learning. Current evidence for sleep learning in humans is inconclusive. To explore conditions under which verbal learning might occur, we hypothesized that peaks of slow waves would be conducive to verbal learning because the peaks define periods of neural excitability. While in slow-wave sleep during a nap, a series of word pairs comprising pseudowords, e.g., "tofer," and actual German words, e.g., "Haus" (house), were played to young German-speaking women and men. When the presentation of the second word of a pair (e.g., "Haus" of "tofer-house") coincided with an ongoing slow-wave peak, the chances increased that a new semantic association between the pair had been formed and retained. Sleep-formed associations translated into awake ones, where they guided forced choices on an implicit memory test. Reactivations of sleep-formed associations were mirrored by brain activation increases measured with fMRI in cortical language areas and the hippocampus, a brain structure critical for relational binding. We infer that implicit relational binding had occurred during peaks of slow oscillations, recruiting a hippocampal-neocortical network comparable to vocabulary learning in the waking state.

Subliminally and Supraliminally Acquired Long-Term Memories Jointly Bias Delayed Decisions.

Common wisdom and scientific evidence suggest that good decisions require conscious deliberation. But growing evidence demonstrates that not only conscious but also unconscious thoughts influence decision-making. Here, we hypothesize that both consciously and unconsciously acquired memories guide decisions. Our experiment measured the influence of subliminally and supraliminally presented information on delayed (30-40 min) decision-making. Participants were presented with subliminal pairs of faces and written occupations for unconscious encoding. Following a delay of 20 min, participants consciously (re-)encoded the same faces now presented supraliminally along with either the same written occupations, occupations congruous to the subliminally presented occupations (same wage-category), or incongruous occupations (opposite wage-category). To measure decision-making, participants viewed the same faces again (with occupations absent) and decided on the putative income of each person: low, low-average, high-average, or high. Participants were encouraged to decide spontaneously and intuitively. Hence, the decision task was an implicit or indirect test of relational memory. If conscious thought alone guided decisions (= H0), supraliminal information should determine decision outcomes independently of the encoded subliminal information. This was, however, not the case. Instead, both unconsciously and consciously encoded memories influenced decisions: identical unconscious and conscious memories exerted the strongest bias on income decisions, while both incongruous and congruous (i.e., non-identical) subliminally and supraliminally formed memories canceled each other out leaving no bias on decisions. Importantly, the increased decision bias following the formation of identical unconscious and conscious memories and the reduced decision bias following to the formation of non-identical memories were determined relative to a control condition, where conscious memory formation alone could influence decisions. In view of the much weaker representational strength of subliminally vs. supraliminally formed memories, their long-lasting impact on decision-making is noteworthy.

Subliminal messages exert long-term effects on decision-making.

Subliminal manipulation is often considered harmless because its effects typically decay within a second. So far, subliminal long-term effects on behavior were only observed in studies which repeatedly presented highly familiar information such as single words. These studies suggest that subliminal messages are only slowly stored and might not be stored at all if they provide novel, unfamiliar information. We speculated that subliminal messages might affect delayed decision-making especially if messages contain several pieces of novel information that must be relationally bound in long-term memory. Relational binding engages the hippocampal memory system, which can rapidly encode and durably store novel relations. Here, we hypothesized that subliminally presented stimulus pairs would be relationally processed influencing the direction of delayed conscious decisions. In experiment 1, subliminal face-occupation pairs affected conscious decisions about the income of these individuals almost half an hour later. In experiment 2, subliminal presentation of vocabulary of a foreign language enabled participants to later decide whether these foreign words are presented with correct or incorrect translations. Subliminal influence did not significantly decay if probed after 25 versus 15 min. This is unprecedented evidence of the longevity and impact of subliminal messages on conscious, rational decision-making.

Hippocampus is place of interaction between unconscious and conscious memories.

Recent evidence suggests that humans can form and later retrieve new semantic relations unconsciously by way of hippocampus-the key structure also recruited for conscious relational (episodic) memory. If the hippocampus subserves both conscious and unconscious relational encoding/retrieval, one would expect the hippocampus to be place of unconscious-conscious interactions during memory retrieval. We tested this hypothesis in an fMRI experiment probing the interaction between the unconscious and conscious retrieval of face-associated information. For the establishment of unconscious relational memories, we presented subliminal (masked) combinations of unfamiliar faces and written occupations ("actor" or "politician"). At test, we presented the former subliminal faces, but now supraliminally, as cues for the reactivation of the unconsciously associated occupations. We hypothesized that unconscious reactivation of the associated occupation-actor or politician-would facilitate or inhibit the subsequent conscious retrieval of a celebrity's occupation, which was also actor or politician. Depending on whether the reactivated unconscious occupation was congruent or incongruent to the celebrity's occupation, we expected either quicker or delayed conscious retrieval process. Conscious retrieval was quicker in the congruent relative to a neutral baseline condition but not delayed in the incongruent condition. fMRI data collected during subliminal face-occupation encoding confirmed previous evidence that the hippocampus was interacting with neocortical storage sites of semantic knowledge to support relational encoding. fMRI data collected at test revealed that the facilitated conscious retrieval was paralleled by deactivations in the hippocampus and neocortical storage sites of semantic knowledge. We assume that the unconscious reactivation has pre-activated overlapping relational representations in the hippocampus reducing the neural effort for conscious retrieval. This finding supports the notion of synergistic interactions between conscious and unconscious relational memories in a common, cohesive hippocampal-neocortical memory space.

Word encoding during sleep is suggested by correlations between word-evoked up-states and post-sleep semantic priming.

To test whether humans can encode words during sleep we played everyday words to men while they were napping and assessed priming from sleep-played words following waking. Words were presented during non-rapid eye movement (NREM) sleep. Priming was assessed using a semantic and a perceptual priming test. These tests measured differences in the processing of words that had been or had not been played during sleep. Synonyms to sleep-played words were the targets in the semantic priming test that tapped the meaning of sleep-played words. All men responded to sleep-played words by producing up-states in their electroencephalogram. Up-states are NREM sleep-specific phases of briefly increased neuronal excitability. The word-evoked up-states might have promoted word processing during sleep. Yet, the mean performance in the priming tests administered following sleep was at chance level, which suggests that participants as a group failed to show priming following sleep. However, performance in the two priming tests was positively correlated to each other and to the magnitude of the word-evoked up-states. Hence, the larger a participant's word-evoked up-states, the larger his perceptual and semantic priming. Those participants who scored high on all variables must have encoded words during sleep. We conclude that some humans are able to encode words during sleep, but more research is needed to pin down the factors that modulate this ability.

Sleep stage II contributes to the consolidation of declarative memories.

Various studies suggest that non-rapid eye movement (NREM) sleep, especially slow-wave sleep (SWS), is vital to the consolidation of declarative memories. However, sleep stage 2 (S2), which is the other NREM sleep stage besides SWS, has gained only little attention. The current study investigated whether S2 during an afternoon nap contributes to the consolidation of declarative memories. Participants learned associations between faces and cities prior to a brief nap. A cued recall test was administered before and following the nap. Spindle, delta and slow oscillation activity was recorded during S2 in the nap following learning and in a control nap. Increases in spindle activity, delta activity, and slow oscillation activity in S2 in the nap following learning compared to the control nap were associated with enhanced retention of face-city associations. Furthermore, spindles tended to occur more frequently during up-states than down-states within slow oscillations during S2 following learning versus S2 of the control nap. These findings suggest that spindles, delta waves, and slow oscillations might promote memory consolidation not only during SWS, as shown earlier, but also during S2.