Long-Horizon Associative Learning Explains Human Sensitivity to Statistical and Network Structures in Auditory Sequences.

Networks are a useful mathematical tool for capturing the complexity of the world. In a previous behavioral study, we showed that human adults were sensitive to the high-level network structure underlying auditory sequences, even when presented with incomplete information. Their performance was best explained by a mathematical model compatible with associative learning principles, based on the integration of the transition probabilities between adjacent and nonadjacent elements with a memory decay. In the present study, we explored the neural correlates of this hypothesis via magnetoencephalography (MEG). Participants (N = 23, 16 females) passively listened to sequences of tones organized in a sparse community network structure comprising two communities. An early difference (∼150 ms) was observed in the brain responses to tone transitions with similar transition probability but occurring either within or between communities. This result implies a rapid and automatic encoding of the sequence structure. Using time-resolved decoding, we estimated the duration and overlap of the representation of each tone. The decoding performance exhibited exponential decay, resulting in a significant overlap between the representations of successive tones. Based on this extended decay profile, we estimated a long-horizon associative learning novelty index for each transition and found a correlation of this measure with the MEG signal. Overall, our study sheds light on the neural mechanisms underlying human sensitivity to network structures and highlights the potential role of Hebbian-like mechanisms in supporting learning at various temporal scales.

Humans parsimoniously represent auditory sequences by pruning and completing the underlying network structure.

Successive auditory inputs are rarely independent, their relationships ranging from local transitions between elements to hierarchical and nested representations. In many situations, humans retrieve these dependencies even from limited datasets. However, this learning at multiple scale levels is poorly understood. Here, we used the formalism proposed by network science to study the representation of local and higher-order structures and their interaction in auditory sequences. We show that human adults exhibited biases in their perception of local transitions between elements, which made them sensitive to high-order network structures such as communities. This behavior is consistent with the creation of a parsimonious simplified model from the evidence they receive, achieved by pruning and completing relationships between network elements. This observation suggests that the brain does not rely on exact memories but on a parsimonious representation of the world. Moreover, this bias can be analytically modeled by a memory/efficiency trade-off. This model correctly accounts for previous findings, including local transition probabilities as well as high-order network structures, unifying sequence learning across scales. We finally propose putative brain implementations of such bias.

Tracking transitional probabilities and segmenting auditory sequences are dissociable processes in adults and neonates.

Since speech is a continuous stream with no systematic boundaries between words, how do pre-verbal infants manage to discover words? A proposed solution is that they might use the transitional probability between adjacent syllables, which drops at word boundaries. Here, we tested the limits of this mechanism by increasing the size of the word-unit to four syllables, and its automaticity by testing asleep neonates. Using markers of statistical learning in neonates' EEG, compared to adult behavioral performances in the same task, we confirmed that statistical learning is automatic enough to be efficient even in sleeping neonates. We also revealed that: (1) Successfully tracking transition probabilities (TP) in a sequence is not sufficient to segment it. (2) Prosodic cues, as subtle as subliminal pauses, enable to recover words segmenting capacities. (3) Adults' and neonates' capacities to segment streams seem remarkably similar despite the difference of maturation and expertise. Finally, we observed that learning increased the overall similarity of neural responses across infants during exposure to the stream, providing a novel neural marker to monitor learning. Thus, from birth, infants are equipped with adult-like tools, allowing them to extract small coherent word-like units from auditory streams, based on the combination of statistical analyses and auditory parsing cues. RESEARCH HIGHLIGHTS: Successfully tracking transitional probabilities in a sequence is not always sufficient to segment it. Word segmentation solely based on transitional probability is limited to bi- or tri-syllabic elements. Prosodic cues, as subtle as subliminal pauses, enable to recover chunking capacities in sleeping neonates and awake adults for quadriplets.

The power of rhythms: how steady-state evoked responses reveal early neurocognitive development.

Electroencephalography (EEG) is a non-invasive and painless recording of cerebral activity, particularly well-suited for studying young infants, allowing the inspection of cerebral responses in a constellation of different ways. Of particular interest for developmental cognitive neuroscientists is the use of rhythmic stimulation, and the analysis of steady-state evoked potentials (SS-EPs) - an approach also known as frequency tagging. In this paper we rely on the existing SS-EP early developmental literature to illustrate the important advantages of SS-EPs for studying the developing brain. We argue that (1) the technique is both objective and predictive: the response is expected at the stimulation frequency (and/or higher harmonics), (2) its high spectral specificity makes the computed responses particularly robust to artifacts, and (3) the technique allows for short and efficient recordings, compatible with infants' limited attentional spans. We additionally provide an overview of some recent inspiring use of the SS-EP technique in adult research, in order to argue that (4) the SS-EP approach can be implemented creatively to target a wide range of cognitive and neural processes. For all these reasons, we expect SS-EPs to play an increasing role in the understanding of early cognitive processes. Finally, we provide practical guidelines for implementing and analyzing SS-EP studies.

Sleeping neonates track transitional probabilities in speech but only retain the first syllable of words.

Extracting statistical regularities from the environment is a primary learning mechanism that might support language acquisition. While it has been shown that infants are sensitive to transition probabilities between syllables in speech, it is still not known what information they encode. Here we used electrophysiology to study how full-term neonates process an artificial language constructed by randomly concatenating four pseudo-words and what information they retain after a few minutes of exposure. Neural entrainment served as a marker of the regularities the brain was tracking during learning. Then in a post-learning phase, evoked-related potentials (ERP) to different triplets explored which information was retained. After two minutes of familiarization with the artificial language, neural entrainment at the word rate emerged, demonstrating rapid learning of the regularities. ERPs in the test phase significantly differed between triplets starting or not with the correct first syllables, but no difference was associated with subsequent violations in transition probabilities. Thus, our results revealed a two-step learning process: neonates segmented the stream based on its statistical regularities, but memory encoding targeted during the word recognition phase entangled the ordinal position of the syllables but was still incomplete at that age.

Automated Pipeline for Infants Continuous EEG (APICE): A flexible pipeline for developmental cognitive studies.

Infant electroencephalography (EEG) presents several challenges compared with adult data: recordings are typically short and heavily contaminated by motion artifacts, and the signal changes throughout development. Traditional data preprocessing pipelines, developed mainly for event-related potential analyses, require manual steps. However, larger datasets make this strategy infeasible. Moreover, new analytical approaches may have different preprocessing requirements. We propose an Automated Pipeline for Infants Continuous EEG (APICE). APICE is fully automated, flexible, and modular. The use of multiple algorithms and adaptive thresholds for artifact detection makes it suitable across age groups and testing procedures. Furthermore, the preprocessing is performed on continuous data, enabling better data recovery and flexibility (i.e., the same preprocessing is usable for different analyzes). Here we describe APICE and validate its performance in terms of data quality and data recovery using two very different infant datasets. Specifically, (1) we show how APICE performs when varying its artifacts rejection sensitivity; (2) we test the effect of different data cleaning methods such as the correction of transient artifacts, Independent Component Analysis, and Denoising Source Separation; and (3) we compare APICE with other available pipelines. APICE uses EEGLAB and compatible custom functions. It is freely available at https://github.com/neurokidslab/eeg_preprocessing, together with example scripts.

Orthogonal neural codes for speech in the infant brain.

Creating invariant representations from an everchanging speech signal is a major challenge for the human brain. Such an ability is particularly crucial for preverbal infants who must discover the phonological, lexical, and syntactic regularities of an extremely inconsistent signal in order to acquire language. Within the visual domain, an efficient neural solution to overcome variability consists in factorizing the input into a reduced set of orthogonal components. Here, we asked whether a similar decomposition strategy is used in early speech perception. Using a 256-channel electroencephalographic system, we recorded the neural responses of 3-mo-old infants to 120 natural consonant-vowel syllables with varying acoustic and phonetic profiles. Using multivariate pattern analyses, we show that syllables are factorized into distinct and orthogonal neural codes for consonants and vowels. Concerning consonants, we further demonstrate the existence of two stages of processing. A first phase is characterized by orthogonal and context-invariant neural codes for the dimensions of manner and place of articulation. Within the second stage, manner and place codes are integrated to recover the identity of the phoneme. We conclude that, despite the paucity of articulatory motor plans and speech production skills, pre-babbling infants are already equipped with a structured combinatorial code for speech analysis, which might account for the rapid pace of language acquisition during the first year.

Remarks on the analysis of steady-state responses: Spurious artifacts introduced by overlapping epochs.

Periodic and stable sensory input can result in rhythmic and stable neural responses, a phenomenon commonly referred to as neural entrainment. Although the use of neural entrainment to investigate the regularities the brain tracks has increased in recent years, the methods used for its quantification are not well-defined in the literature. Here we argue that some strategies used in previous papers, are inadequate for the study of steady-state response, and lead to methodological artefacts. The aim of this commentary is to discuss these articles and to propose alternative measures of neural entrainment. Specifically, we applied four possible alternatives and two epoching approaches reported in the literature to quantify neural entrainment on simulated datasets. Our results demonstrate that overlapping epochs, as used in the original Batterink and colleagues articles, inevitably lead to a methodological artefact at the frequency corresponding to the overlap. We therefore strongly discourage this approach and encourage the re-analysis of data based on overlapping epochs. Additionally, we argue that the use of time-frequency decomposition to compute phase coherence at low frequencies to reveal neural entrainment is not optimal.

Prosody facilitates learning the word order in a new language.

One of the prominent ideas developed by Jacques Mehler and his colleagues was that perceptual tuning, present from birth on, enables infants, and language learners in general, to extract regularities from speech input. Here we discuss language learners'' ability to extract basic word order (VO or OV) structure from prosodic regularities in a language. The two are closely related: in phonological phrases of VO languages, the most prominent word is the rightmost one, and in OV languages, it is the leftmost one. In speech, this prominence is realized as extended duration, or as elevated pitch, sometimes combined with changes in intensity. When learning the first (L1) or the second language (L2), exposure to relevant rhythmic structure elicits implicit learning about syntactic structure, including the basic word order. However, it remains unclear whether triggering the learning process requires a certain level of familiarity with the relevant rhythm. It is moreover unknown whether prosodic information can help L2 learners to extract and learn the vocabulary of a new language. We tested Spanish- and Italian-speaking adults' ability to learn words from an artificial language with either non-native OV or native VO word order. The results show that learners used prosodic information to identify the most prominent words in short utterances when the artificial language was similar to the native language, with duration-based prominence in prosody and a VO word order. In contrast, when the artificial language had a non-native prominence marked by pitch alternations and an OV word order, prominent words were learned only after a three-day exposure to the relevant rhythmic structure. Thus, for adult L2 learners, only repeated exposure to the relevant prosody elicited learning new words from an unknown language with non-native prosodic marking, indicating that, with familiarity, prosodic cues can facilitate learning in L2.

Evidence of ordinal position encoding of sequences extracted from continuous speech.

Infants' capacity to extract statistical regularities from sequential information is impressive and well documented. However, statistical learning's underlying mechanism remains mostly unknown, and its role in language acquisition is still under debate. To shed light on these issues, here we address the question of which information human subjects extract and encode after familiarisation with a continuous sequence of stimuli and its dependence on the type of segmentation cues and on the stimuli modality. Specifically, we investigate whether adults and 5-month-old infants learn the syllables' co-occurrence in the stream or generate a representation of the Words that include syllables' ordinal position. We test if subtle pauses signalling word boundaries change the encoding and, in adults, if it varies across modalities. In six behavioural experiments, we show that: (i) Adults and infants learn the streams' statistical structure. (ii) Ordinal encoding emerges in the auditory modality, and pauses enhanced it. However, (iii) ordinal encoding seems to depend on the learning stage and not on pauses marking Words' edges. Interestingly, (iv) for visual presentation of orthographic syllables, we do not find evidence of ordinal encoding in adults. Our results support the emergence, in the auditory modality, of a Word representation where its constituents are associated with an ordinal position at play already early in life, bringing new insights into speech processing and language acquisition. Additionally, we successfully use for the first time pupillometry in an infant segmentation task.

Newborns are sensitive to multiple cues for word segmentation in continuous speech.

Before infants can learn words, they must identify those words in continuous speech. Yet, the speech signal lacks obvious boundary markers, which poses a potential problem for language acquisition (Swingley, Philos Trans R Soc Lond. Series B, Biol Sci 364(1536), 3617-3632, 2009). By the middle of the first year, infants seem to have solved this problem (Bergelson & Swingley, Proc Natl Acad Sci 109(9), 3253-3258, 2012; Jusczyk & Aslin, Cogn Psychol 29, 1-23, 1995), but it is unknown if segmentation abilities are present from birth, or if they only emerge after sufficient language exposure and/or brain maturation. Here, in two independent experiments, we looked at two cues known to be crucial for the segmentation of human speech: the computation of statistical co-occurrences between syllables and the use of the language's prosody. After a brief familiarization of about 3 min with continuous speech, using functional near-infrared spectroscopy, neonates showed differential brain responses on a recognition test to words that violated either the statistical (Experiment 1) or prosodic (Experiment 2) boundaries of the familiarization, compared to words that conformed to those boundaries. Importantly, word recognition in Experiment 2 occurred even in the absence of prosodic information at test, meaning that newborns encoded the phonological content independently of its prosody. These data indicate that humans are born with operational language processing and memory capacities and can use at least two types of cues to segment otherwise continuous speech, a key first step in language acquisition.

In vivo measure of neonate brain optical properties and hemodynamic parameters by time-domain near-infrared spectroscopy.

By exploiting a multichannel portable instrument for time-domain near-infrared spectroscopy (TD-NIRS), we characterized healthy neonates' brains in term of optical properties and hemodynamic parameters. In particular, we assessed the absolute values of the absorption and reduced scattering coefficients at two wavelengths, together with oxy-, deoxy- and total hemoglobin concentrations, and the blood oxygen saturation of the neonates' brains. In this study, 33 healthy full-term neonates were tested, obtaining the following median values: 0.28 and [Formula: see text] for [Formula: see text] at 690 and 820 nm, respectively; 5.8 and [Formula: see text] for [Formula: see text] at 690 and 820 nm, respectively; [Formula: see text] for [Formula: see text]; [Formula: see text] for [Formula: see text]; [Formula: see text] for [Formula: see text]; 72% for [Formula: see text]. In general, the agreement of these values with the sparse existing literature appears not always consistent. These findings demonstrate the first measurements of optical properties of the healthy neonate brain using TD-NIRS and show the need for clarification of optical properties across methods and populations.

Daily Variation of UV-induced Erythema and the Action of Solar Filters.

UV rays may cause several degrees of skin damage, which makes sunscreen research necessary. In addition, skin sensitivity shows daily variations, which can interfere in the detection of the efficacy of the filters. Here, we studied the UV-induced erythema in hairless rats at two times of the day (light and darkness) using a colorimeter method. The effect of an emulsion with solar filters with or without melatonin was also assayed. Results indicate that the value of a* (from CIELAB color space values L* a* b) was the most useful variable to evaluate the erythema. However, at the UV intensity used, erythema was only detected when irradiation was carried out during the activity phase of the animal, enabling the detection of the protective action of the sunscreen at this time. Thus, daily variations in skin sensitivity have been demonstrated and should be taken into account in dermatological research.

Melatonin pharmacokinetics after transdermal administration changes according to the time of the day.

Melatonin is a neurohormone with multiple and different actions, such as chronobiotic or antioxidant. Melatonin is usually orally administered, but dermal administration is also useful in dermatological diseases or as adjuvant to certain skin treatments. Here, we studied the variability of the pharmacokinetics of melatonin and its metabolite AFMK, when melatonin is transdermally administered to Hairless rat at two different times of day (Zeitgeber Time 4 (ZT4) and ZT16). Moreover, in order to obtain the bioavailability, kinetics after intravenous administration was also studied. In addition, a permeation study was carried out, at both ZTs, to test the amount of melatonin retained in the skin after transdermal administration. Results showed that pharmacokinetic parameters of melatonin administered exogenously depended on the time of the day. When intravenous data were fitted to a compartmental model, the extrapolated plasma concentration at time 0 and the area under the curve were higher at ZT4, while clearance, volumes of central and peripheral compartments and volume of distribution at the steady state were higher at ZT16. Transdermal administration was best fitted to a one-compartment model and tmax, half-life of absorption and area under the curve showed higher values at ZT4, while the absorption rate and constant of absorption were higher at ZT16. AFMK was detected in all cases, but no differences between the two ZTs were observed. Transdermal administration showed better bioavailability also at ZT4. Results indicate that time of day is a variable that should be taken into account when melatonin is transdermally administered.

Melatonin Delivery: Transdermal and Transbuccal Evaluation in Different Vehicles.

PURPOSE: Melatonin (MLT) could be candidate drug for treatment of several diseases because of its high antioxidant and anticarcinogenic activity and its important biological roles. The aim of this study was to assess the influence of different vehicles on the permeation of MLT through buccal and skin tissues. METHODS: Formulations were characterized in terms of rheology, drug release and permeation through human skin as well as porcine buccal mucosa. Irradiation experiments were also performed. RESULTS: The lowest amount of MLT released was from oral adhesive paste Orabase® (OB) and the highest from the emulsion system Montanov® 68 (M68). Skin permeation revealed high pattern for Carbopol® 940 (C940) and M68, and poor for poloxamer 407 (P407) and Pluronic® lecithin organogel (PLO). Statistical differences of MLT remaining in skin between M68 vs C940 (p < 0.05) and M68 vs PLO (p < 0.05) were observed. Transmucosal results showed that sodium carboxymethylcellulose (NaCMC) was the best and OB the worst vehicle. P407 and PLO followed similar behaviour. Photostability studies revealed high percentage of degradation of MLT in solution which was also similar when was loaded in OB. The rest of formulations showed low rates of degradation. CONCLUSIONS: C940 or M68 and NaCMC can be proposed as formulations for a potential systemic effect of MLT by skin and buccal mucosa routes, respectively. However, if the intended objective is to obtain local action in the skin and buccal mucosa, the proposed formulations are M68 or P407 and PLO.

On the edge of language acquisition: inherent constraints on encoding multisyllabic sequences in the neonate brain.

To understand language, humans must encode information from rapid, sequential streams of syllables - tracking their order and organizing them into words, phrases, and sentences. We used Near-Infrared Spectroscopy (NIRS) to determine whether human neonates are born with the capacity to track the positions of syllables in multisyllabic sequences. After familiarization with a six-syllable sequence, the neonate brain responded to the change (as shown by an increase in oxy-hemoglobin) when the two edge syllables switched positions but not when two middle syllables switched positions (Experiment 1), indicating that they encoded the syllables at the edges of sequences better than those in the middle. Moreover, when a 25 ms pause was inserted between the middle syllables as a segmentation cue, neonates' brains were sensitive to the change (Experiment 2), indicating that subtle cues in speech can signal a boundary, with enhanced encoding of the syllables located at the edges of that boundary. These findings suggest that neonates' brains can encode information from multisyllabic sequences and that this encoding is constrained. Moreover, subtle segmentation cues in a sequence of syllables provide a mechanism with which to accurately encode positional information from longer sequences. Tracking the order of syllables is necessary to understand language and our results suggest that the foundations for this encoding are present at birth.

Extinction interferes with the retrieval of visuomotor memories through a mechanism involving the sensorimotor cortex.

Savings is a fundamental property of learning. In motor adaptation, it refers to the improvement in learning observed when adaptation to a perturbation A (A1) is followed by re-adaptation to the same perturbation (A2). A common procedure to equate the initial level of error across sessions consists of restoring native sensorimotor coordinates by inserting null--unperturbed--trials (N) just before re-adaptation (washout). Here, we hypothesized that the washout is not innocuous but interferes with the expression of the new memory at recall. To assess this possibility, we measured savings following the A1NA2 protocol, where A was a 40° visual rotation. In Experiment 1, we increased the time window between N and A2 from 1 min to 24 h. This manipulation increased the amount of savings during middle to late phases of adaptation, suggesting that N interfered with the retrieval of A. In Experiment 2, we used repetitive TMS to evaluate if this interference was partly mediated by the sensorimotor cortex (SM). We conclude that the washout does not just restore the unperturbed sensorimotor coordinates, but inhibits the expression of the recently acquired visuomotor map through a mechanism involving SM. Our results resemble the phenomenon of extinction in classical conditioning.

Circadian rhythms on skin function of hairless rats: light and thermic influences.

Circadian rhythms are present in most functions of living beings. We have demonstrated the presence of circadian rhythms in skin variables (transepidermal water loss, TEWL; stratum corneum hydration, SCH; and skin temperature) in hairless rats under different environmental conditions of light and temperature. Circadian rhythms in TEWL and SCH showed mean amplitudes of about 20% and 14% around the mean, respectively, and appeared under light-dark cycles as well as under constant darkness. Environmental temperature was able to override TEWL, but not SCH rhythm, evidencing the dependency of TEWL on the temperature. Mean daily values of TEWL and SCH, and also the amplitude of TEWL rhythm, increased with the age of the animal. Under constant light, situation that induces arrhythmicity in rats, SCH and TEWL were inversely correlated. The results suggest the importance to take into account the functional skin rhythms in research in dermatological sciences.