Optimized single-cell RNA sequencing protocol to study early genome activation in mammalian preimplantation development.

Here, we present a modification of single-cell tagged reverse transcription protocol to study gene expression on a single-cell level or with limited RNA input. We describe different enzymes for reverse transcription and cDNA amplification, modified lysis buffer, and additional clean-up steps before cDNA amplification. We also detail an optimized single-cell RNA sequencing method for handpicked single cells, or tens to hundreds of cells, as input material to study mammalian preimplantation development. For complete details on the use and execution of this protocol, please refer to Ezer et al.1.

The neuroscience of music - towards ecological validity.

Studies in the neuroscience of music gained momentum in the 1990s as an integrated part of the well-controlled experimental research tradition. However, during the past two decades, these studies have moved toward more naturalistic, ecologically valid paradigms. Here, I introduce this move in three frameworks: (i) sound stimulation and empirical paradigms, (ii) study participants, and (iii) methods and contexts of data acquisition. I wish to provide a narrative historical overview of the development of the field and, in parallel, to stimulate innovative thinking to further advance the ecological validity of the studies without overlooking experimental rigor.

Comprehensive characterization of the embryonic factor LEUTX.

The paired-like homeobox transcription factor LEUTX is expressed in human preimplantation embryos between the 4- and 8-cell stages, and then silenced in somatic tissues. To characterize the function of LEUTX, we performed a multiomic characterization of LEUTX using two proteomics methods and three genome-wide sequencing approaches. Our results show that LEUTX stably interacts with the EP300 and CBP histone acetyltransferases through its 9 amino acid transactivation domain (9aaTAD), as mutation of this domain abolishes the interactions. LEUTX targets genomic cis-regulatory sequences that overlap with repetitive elements, and through these elements it is suggested to regulate the expression of its downstream genes. We find LEUTX to be a transcriptional activator, upregulating several genes linked to preimplantation development as well as 8-cell-like markers, such as DPPA3 and ZNF280A. Our results support a role for LEUTX in preimplantation development as an enhancer binding protein and as a potent transcriptional activator.

Mismatch negativity-stimulation paradigms in past and in future.

Mismatch negativity (MMN) studies were initiated as part of a well-controlled experimental research tradition with the aim to identify some key principles of auditory processing and memory. During the past two decades, empirical paradigms have moved toward more ecologically valid ones while retaining rigid experimental control. In this paper, I will introduce this development of MMN stimulation paradigms starting from the paradigms used in basic science and then moving to paradigms that have been particularly relevant for studies on music learning and musical expertise. Via these historical and thematic perspectives, I wish to stimulate paradigm development further to meet the demands of naturalistic ecologically valid studies also when using MMN in the context of event-related potential technique that necessarily requires averaging across several stimulus presentations.

Children's inhibition skills are associated with their P3a latency-results from an exploratory study.

BACKGROUND: The P3a response is thought to reflect involuntary orienting to an unexpected stimulus and has been connected with set-shifting and inhibition in some studies. In our exploratory study, we investigated if the amplitude and the latency of the P3a response were associated with the performance in a modified flanker task measuring inhibition and set-shifting in 10-year-old children (N = 42). Children participated in electroencephalography (EEG) measurement with an auditory multifeature paradigm including standard, deviating, and novel sounds. In addition, they performed a separate flanker task requiring inhibition and set-shifting skills. RESULTS: The P3a latencies for deviant sounds were associated with the reaction time reflecting inhibition: the shorter the response latencies were, the faster the reaction time was. The P3a latencies for novel sounds were not linked to the reaction times reflecting either inhibition or set-shifting. In addition, the magnitude of the P3a response was not associated with the performance in the flanker task. CONCLUSIONS: Our results suggest that P3a response latency and reaction speed reflecting inhibitory skills are based on shared neural mechanism. Thus, the present study brings new insight to the field investigating the associations between behavior and its neural indices.

Identifying Personality Characteristics and Indicators of Psychological Well-Being Associated With Attrition in the Motivation Makes the Move! Physical Activity Intervention: Randomized Technology-Supported Trial.

BACKGROUND: Data attrition has been a common problem in longitudinal lifestyle interventions. The contributors to attrition in technology-supported physical activity interventions have not been thoroughly studied. OBJECTIVE: The present study examined the roles of personality characteristics and indicators of psychological well-being in data attrition within a technology-supported, longitudinal intervention study with overweight adults. METHODS: Participants (N=89) were adults from the Motivation Makes the Move! intervention study. Data attrition was studied after a 3-month follow-up. Participants' personality characteristics were studied using the Short Five self-report questionnaire. Psychological well-being indicators were assessed with the RAND 36-item health survey, Positive and Negative Affect Schedule, and Beck Depression Inventory. Logistic regression analyses were conducted to assess the risk of discontinuing the study. The analyses were adjusted for sex, age, study group, and educational status. RESULTS: At the 3-month follow-up, 65 of 89 participants (73% of the initial sample) had continued in the study. Participants' personality characteristics and indicators of psychological well-being were not associated with the risk of dropping out of the study (all P values >.05). The results remained the same after covariate controls. CONCLUSIONS: Participant attrition was not attributable to personality characteristics or psychological well-being in the Motivation Makes the Move! study conducted with overweight adults. As attrition remains a challenge within longitudinal, technology-supported lifestyle interventions, attention should be paid to the potentially dynamic natures of personality and psychological well-being, as well as other elements beyond these. TRIAL REGISTRATION: ClinicalTrials.gov NCT02686502; https://clinicaltrials.gov/ct2/show/NCT02686502.

Music-based interventions in community settings: Navigating the tension between rigor and ecological validity.

Empirical research of community-based music interventions has advanced to investigate the individual, social, and educational implications of arts-for-wellbeing practices. Here, we present the motivations, aims, hypotheses, and implications of this complex field of inquiry. We describe examples of recent large-scale investigations to reflect on the major methodological challenges. Community-based music interventions strike a balance between the empirical rigor of clinical trials and the demands of ecological validity. We argue that this balance should be viewed as an asset rather than a mere pragmatic compromise. We also offer some perspectives on best-practice models for effectively engaging in this type of work.

Transient DUX4 expression in human embryonic stem cells induces blastomere-like expression program that is marked by SLC34A2.

Embryonic genome activation (EGA) is critical for embryonic development. However, our understanding of the regulatory mechanisms of human EGA is still incomplete. Human embryonic stem cells (hESCs) are an established model for studying developmental processes, but they resemble epiblast and are sub-optimal for modeling EGA. DUX4 regulates human EGA by inducing cleavage-stage-specific genes, while it also induces cell death. We report here that a short-pulsed expression of DUX4 in primed hESCs activates an EGA-like gene expression program in up to 17% of the cells, retaining cell viability. These DUX4-induced cells resembled eight-cell stage blastomeres and were named induced blastomere-like (iBM) cells. The iBM cells showed marked reduction of POU5F1 protein, as previously observed in mouse two-cell-like cells. Finally, the iBM cells were successfully enriched using an antibody against NaPi2b (SLC34A2), which is expressed in human blastomeres. The iBM cells provide an improved model system to study human EGA transcriptome.

Inter-brain synchronization occurs without physical co-presence during cooperative online gaming.

Inter-brain synchronization during social interaction has been linked with several positive phenomena, including closeness, cooperation, prosociality, and team performance. However, the temporal dynamics of inter-brain synchronization during collaboration are not yet fully understood. Furthermore, with collaboration increasingly happening online, the dependence of inter-brain phase synchronization of oscillatory activity on physical presence is an important but understudied question. In this study, physically isolated participants performed a collaborative coordination task in the form of a cooperative multiplayer game. We measured EEG from 42 subjects working together as pairs in the task. During the measurement, the only interaction between the participants happened through on-screen movement of a racing car, controlled by button presses of both participants working with distinct roles, either controlling the speed or the direction of the car. Pairs working together in the task were found to have elevated neural coupling in the alpha, beta, and gamma frequency bands, compared to performance matched false pairs. Higher gamma synchrony was associated with better momentary performance within dyads and higher alpha synchrony was associated with better mean performance across dyads. These results are in line with previous findings of increased inter-brain synchrony during interaction, and show that phase synchronization of oscillatory activity occurs during online real-time joint coordination without any physical co-presence or video and audio connection. Synchrony decreased during a playing session, but was found to be higher during the second session compared to the first. The novel paradigm, developed for the measurement of real-time collaborative performance, demonstrates that changes in inter-brain EEG phase synchrony can be observed continuously during interaction.

Benefits of choir singing on complex auditory encoding in the aging brain: An ERP study.

Aging is accompanied by difficulties in auditory information processing, especially in more complex sound environments. Choir singing requires efficient processing of multiple sound features and could, therefore, mitigate the detrimental effects of aging on complex auditory encoding. We recorded auditory event-related potentials during passive listening of sounds in healthy older adult (≥ 60 years) choir singers and nonsinger controls. We conducted a complex oddball condition involving encoding of abstract regularities in combinations of pitch and location features, as well as in two simple oddball conditions, in which only either the pitch or spatial location of the sounds was varied. We analyzed change-related mismatch negativity (MMN) and obligatory P1 and N1 responses in each condition. In the complex condition, the choir singers showed a larger MMN than the controls, which also correlated with better performance in a verbal fluency test. In the simple pitch and location conditions, the choir singers had smaller N1 responses compared to the control subjects, whereas the MMN responses did not differ between groups. These results suggest that regular choir singing is associated both with more enhanced encoding of complex auditory regularities and more effective adaptation to simple sound features.

Exploring body consciousness of dancers, athletes, and lightly physically active adults.

Body consciousness is associated with kinetic skills and various aspects of wellbeing. Physical activities have been shown to contribute to the development of body consciousness. Methodological studies are needed in improving the assessment of body consciousness in adults with distinct physical activity backgrounds. This study (1) examined whether dancers, athletes, and lightly physically active individuals differed regarding the level of their body consciousness, and (2) evaluated the usability of different methods in assessing body consciousness. Fifty-seven healthy adults (aged 20-37) were included in the study. Three experimental methods (aperture task, endpoint matching, and posture copying) and two self-report questionnaires (the Private Body Consciousness Scale, PBCS, and the Body Awareness Questionnaire, BAQ) were used in assessing body consciousness. Athletes outperformed the lightly physically active participants in the posture copying task with the aid of vision when copying leg postures. Dancers performed better than the athletes without the aid of vision when their back and upper body were involved, and better than the lightly active participants when copying leg postures. Dancers and athletes had higher self-reported cognitive and perceptual knowledge of their body than lightly physically active participants. To examine the role of different physical activities in developing body consciousness, experimental methods involving the use of the whole body might be most suitable. Subjective measures may provide complementary evidence for experimental testing.

Musical perceptual skills, but not neural auditory processing, are associated with better reading ability in childhood.

Musical activities have been suggested to be beneficial for language development in childhood. Randomised controlled trials using music have indicated that musical interventions can be used to support language skills in children with developmental language difficulties. However, it is not entirely clear how beneficial music activities are for normally developing children or how the effects mediated via music are transmitted. To investigate these questions, the present study used structural equation models to assess how musical training, perceptual musical skills, and auditory processing in the brain are associated with reading proficiency and each other. Perceptual musical skills were assessed using musicality tests while auditory processing in the brain was measured using mismatch negativity responses to pitch, duration, and phoneme length contrasts. Our participants were a community sample of 64 8-11-year-old typically developing children with and without musical training, recruited from four classes in four elementary schools in Finland. Approximately half of children had music as a hobby. Our results suggest that performance in tests of musical perceptual skills is directly linked with reading proficiency instead of being mediated via auditory processing in the brain. Auditory processing in the brain in itself seems not to be strongly linked with reading proficiency in these children. Our results support the view that musical perceptual skills are associated with reading skills regardless of musical training.

Brain oscillation recordings of the audience in a live concert-like setting.

There are only a few previous EEG studies that were conducted while the audience is listening to live music. However, in laboratory settings using music recordings, EEG frequency bands theta and alpha are connected to music improvisation and creativity. Here, we measured EEG of the audience in a concert-like setting outside the laboratory and compared the theta and alpha power evoked by partly improvised versus regularly performed familiar versus unfamiliar live classical music. To this end, partly improvised and regular versions of pieces by Bach (familiar) and Melartin (unfamiliar) were performed live by a chamber trio. EEG data from left and right frontal and central regions of interest were analysed to define theta and alpha power during each performance. After the performances, the participants rated how improvised and attractive each of the performances were. They also gave their affective ratings before and after each performance. We found that theta power was enhanced during the familiar improvised Bach piece and the unfamiliar improvised Melartin piece when compared with the performance of the same piece performed in a regular manner. Alpha power was not modulated by manner of performance or by familiarity of the piece. Listeners rated partly improvised performances of a familiar Bach and unfamiliar Melartin piece as more improvisatory and innovative than the regular performances. They also indicated more joy and less sadness after listening to the unfamiliar improvised piece of Melartin and less fearful and more enthusiastic after listening to the regular version of Melartin than before listening. Thus, according to our results, it is possible to study listeners' brain functions with EEG during live music performances outside the laboratory, with theta activity reflecting the presence of improvisation in the performances.

Improved Auditory Function Caused by Music Versus Foreign Language Training at School Age: Is There a Difference?

In adults, music and speech share many neurocognitive functions, but how do they interact in a developing brain? We compared the effects of music and foreign language training on auditory neurocognition in Chinese children aged 8-11 years. We delivered group-based training programs in music and foreign language using a randomized controlled trial. A passive control group was also included. Before and after these year-long extracurricular programs, auditory event-related potentials were recorded (n = 123 and 85 before and after the program, respectively). Through these recordings, we probed early auditory predictive brain processes. To our surprise, the language program facilitated the children's early auditory predictive brain processes significantly more than did the music program. This facilitation was most evident in pitch encoding when the experimental paradigm was musically relevant. When these processes were probed by a paradigm more focused on basic sound features, we found early predictive pitch encoding to be facilitated by music training. Thus, a foreign language program is able to foster auditory and music neurocognition, at least in tonal language speakers, in a manner comparable to that by a music program. Our results support the tight coupling of musical and linguistic brain functions also in the developing brain.

Psychological and Physiological Signatures of Music Listening in Different Listening Environments-An Exploratory Study.

We compared music emotion ratings and their physiological correlates when the participants listened to music at home and in the laboratory. We hypothesized that music emotions are stronger in a familiar environment, that is, at home. Participants listened to their self-selected favorite and neutral music excerpts at home and in the laboratory for 10 min in each environment. They completed the questionnaires about their emotional states and gave saliva samples for the analyses of the stress hormone cortisol. We found that in the context of music listening, the participants' emotion ratings differed between home and the laboratory. Furthermore, the cortisol levels were generally lower at home than in the laboratory and decreased after music listening at home and in the laboratory. However, the modulatory effects of music listening on cortisol levels did not differ between the home and the laboratory. Our exploratory multimethodological data offer novel insight about the psychological and physiological consequences of music listening. These data reveal the sensitivity of the current research methods to investigate human emotions in various contexts without excluding the use of laboratory environment in investigating them.

Faster maturation of selective attention in musically trained children and adolescents: Converging behavioral and event-related potential evidence.

Previous work suggests that musical training in childhood is associated with enhanced executive functions. However, it is unknown whether this advantage extends to selective attention-another central aspect of executive control. We recorded a well-established event-related potential (ERP) marker of distraction, the P3a, during an audio-visual task to investigate the maturation of selective attention in musically trained children and adolescents aged 10-17 years and a control group of untrained peers. The task required categorization of visual stimuli, while a sequence of standard sounds and distracting novel sounds were presented in the background. The music group outperformed the control group in the categorization task and the younger children in the music group showed a smaller P3a to the distracting novel sounds than their peers in the control group. Also, a negative response elicited by the novel sounds in the N1/MMN time range (~150-200 ms) was smaller in the music group. These results indicate that the music group was less easily distracted by the task-irrelevant sound stimulation and gated the neural processing of the novel sounds more efficiently than the control group. Furthermore, we replicated our previous finding that, relative to the control group, the musically trained children and adolescents performed faster in standardized tests for inhibition and set shifting. These results provide novel converging behavioral and electrophysiological evidence from a cross-modal paradigm for accelerated maturation of selective attention in musically trained children and adolescents and corroborate the association between musical training and enhanced inhibition and set shifting.

Effects of Music Training on the Auditory Working Memory of Chinese-Speaking School-Aged Children: A Longitudinal Intervention Study.

Music expertise is known to be beneficial for cognitive function and development. In this study, we conducted 1-year music training for school children (n = 123; 7-11 years of age before training) in China. The children were assigned to music or second-language after-class training groups. A passive control group was included. We aimed to investigate whether music training could facilitate working memory (WM) development compared to second-language training and no training. Before and after the training, auditory WM was measured via a digit span (DS) task, together with the vocabulary and block tests of the Wechsler Intelligence Scale for Child IV (WISC-IV). The results of the DS task revealed superior development in the music group compared to the other groups. However, further analysis of DS forward and backward tasks indicated that the performance of the three training/non-training groups only differed significantly in DS backward scores, but not in the DS forward scores. We conclude that music training may benefit the central executive system of WM, as reflected by the DS backward task.

Vocal music enhances memory and language recovery after stroke: pooled results from two RCTs.

OBJECTIVE: Previous studies suggest that daily music listening can aid stroke recovery, but little is known about the stimulus-dependent and neural mechanisms driving this effect. Building on neuroimaging evidence that vocal music engages extensive and bilateral networks in the brain, we sought to determine if it would be more effective for enhancing cognitive and language recovery and neuroplasticity than instrumental music or speech after stroke. METHODS: Using data pooled from two single-blind randomized controlled trials in stroke patients (N = 83), we compared the effects of daily listening to self-selected vocal music, instrumental music, and audiobooks during the first 3 poststroke months. Outcome measures comprised neuropsychological tests of verbal memory (primary outcome), language, and attention and a mood questionnaire performed at acute, 3-month, and 6-month stages and structural and functional MRI at acute and 6-month stages. RESULTS: Listening to vocal music enhanced verbal memory recovery more than instrumental music or audiobooks and language recovery more than audiobooks, especially in aphasic patients. Voxel-based morphometry and resting-state and task-based fMRI results showed that vocal music listening selectively increased gray matter volume in left temporal areas and functional connectivity in the default mode network. INTERPRETATION: Vocal music listening is an effective and easily applicable tool to support cognitive recovery after stroke as well as to enhance early language recovery in aphasia. The rehabilitative effects of vocal music are driven by both structural and functional plasticity changes in temporoparietal networks crucial for emotional processing, language, and memory.