MYC Recruits SPT5 to RNA Polymerase II to Promote Processive Transcription Elongation.

The MYC oncoprotein binds to promoter-proximal regions of virtually all transcribed genes and enhances RNA polymerase II (Pol II) function, but its precise mode of action is poorly understood. Using mass spectrometry of both MYC and Pol II complexes, we show here that MYC controls the assembly of Pol II with a small set of transcription elongation factors that includes SPT5, a subunit of the elongation factor DSIF. MYC directly binds SPT5, recruits SPT5 to promoters, and enables the CDK7-dependent transfer of SPT5 onto Pol II. Consistent with known functions of SPT5, MYC is required for fast and processive transcription elongation. Intriguingly, the high levels of MYC that are expressed in tumors sequester SPT5 into non-functional complexes, thereby decreasing the expression of growth-suppressive genes. Altogether, these results argue that MYC controls the productive assembly of processive Pol II elongation complexes and provide insight into how oncogenic levels of MYC permit uncontrolled cellular growth.

Repression of Divergent Noncoding Transcription by a Sequence-Specific Transcription Factor.

Many active eukaryotic gene promoters exhibit divergent noncoding transcription, but the mechanisms restricting expression of these transcripts are not well understood. Here, we demonstrate how a sequence-specific transcription factor represses divergent noncoding transcription at highly expressed genes in yeast. We find that depletion of the transcription factor Rap1 induces noncoding transcription in a large fraction of Rap1-regulated gene promoters. Specifically, Rap1 prevents transcription initiation at cryptic promoters near its binding sites, which is uncoupled from transcription regulation in the protein-coding direction. We further provide evidence that Rap1 acts independently of previously described chromatin-based mechanisms to repress cryptic or divergent transcription. Finally, we show that divergent transcription in the absence of Rap1 is elicited by the RSC chromatin remodeler. We propose that a sequence-specific transcription factor limits access of basal transcription machinery to regulatory elements and adjacent sequences that act as divergent cryptic promoters, thereby providing directionality toward productive transcription.

General Regulatory Factors Control the Fidelity of Transcription by Restricting Non-coding and Ectopic Initiation.

The fidelity of transcription initiation is essential for accurate gene expression, but the determinants of start site selection are not fully understood. Rap1 and other general regulatory factors (GRFs) control the expression of many genes in yeast. We show that depletion of these factors induces widespread ectopic transcription initiation within promoters. This generates many novel non-coding RNAs and transcript isoforms with diverse stability, drastically altering the coding potential of the transcriptome. Ectopic transcription initiation strongly correlates with altered nucleosome positioning. We provide evidence that Rap1 can suppress ectopic initiation by a "place-holder" mechanism whereby it physically occludes inappropriate sites for pre-initiation complex formation. These results reveal an essential role for GRFs in the fidelity of transcription initiation and in the suppression of pervasive transcription, profoundly redefining current models for their function. They have important implications for the mechanism of transcription initiation and the control of gene expression.

Large-Area Perovskite Nanocrystal Metasurfaces for Direction-Tunable Lasing.

Perovskite nanocrystals (PNCs) are attractive emissive materials for developing compact lasers. However, manipulation of PNC laser directionality has been difficult, which limits their usage in photonic devices that require on-demand tunability. Here we demonstrate PNC metasurface lasers with engineered emission angles. We fabricated millimeter-scale CsPbBr3 PNC metasurfaces using an all-solution-processing technique based on soft nanoimprinting lithography. By designing band-edge photonic modes at the high-symmetry X point of the reciprocal lattice, we achieved four linearly polarized lasing beams along a polar angle of ∼30° under optical pumping. The device architecture further allows tuning of the lasing emission angles to 0° and ∼50°, respectively, by adjusting the PNC thickness to shift other high-symmetry points (Γ and M) to the PNC emission wavelength range. Our laser design strategies offer prospects for applications in directional optical antennas and detectors, 3D laser projection displays, and multichannel visible light communication.

Giant Photoluminescence Enhancement of Monolayer WSe2 Using a Plasmonic Nanocavity with On-Demand Resonance.

Monolayer transition metal dichalcogenides (TMDs) are considered promising building blocks for next-generation photonic and optoelectronic devices, owing to their fascinating optical properties. However, their inherent weak light absorption and low quantum yield severely hinder their practical applications. Here, we report up to 18000-fold photoluminescence (PL) enhancement in a monolayer WSe2-coupled plasmonic nanocavity. A spectroscopy-assisted nanomanipulation technique enables the assembly of a nanocavity with customizable resonances to simultaneously enhance the excitation and emission processes. In particular, precise control over the magnetic cavity mode facilitates spectral and spatial overlap with the exciton, resulting in plasmon-exciton intermediate coupling that approaches the maximum emission rate in the hybrid system. Meanwhile, the cavity mode exhibits high radiation directivity, which overwhelmingly directs surface-normal PL emission and leads to a 17-fold increase in the collection efficiency. Our approach opens up a new avenue to enhance the PL intensity of monolayer TMDs, facilitating their implementation in highly efficient optoelectronic devices.

Unidirectional Lasing from Mirror-Coupled Dielectric Lattices.

This paper reports how a hybrid system composed of transparent dielectric lattices over a metal mirror can produce high-quality lattice resonances for unidirectional lasing. The enhanced electromagnetic fields are concentrated in the cladding of the periodic dielectric structures and away from the metal. Based on a mirror-image model, we reveal that such high-quality lattice resonances are governed by bound states in the continuum resulting from destructive interference. Using hexagonal arrays of titanium dioxide nanoparticles on a silica-coated silver mirror, we observed lattice resonances with quality factors of up to 2750 in the visible regime. With the lattice resonances as optical feedback and dye solution as the gain medium, we demonstrated unidirectional lasing under optical pumping, where the array size was down to 100 µm × 100 µm. Our scheme can be extended to other spectral regimes to simultaneously achieve strongly enhanced surface fields and high quality factors.

Lead/Lag directionality is not generally equivalent to causality in nonlinear systems: Comparison of phase slope index and conditional mutual information.

Applications of causal techniques to neural time series have increased extensively over last decades, including a wide and diverse family of methods focusing on electroencephalogram (EEG) analysis. Besides connectivity inferred in defined frequency bands, there is a growing interest in the analysis of cross-frequency interactions, in particular phase and amplitude coupling and directionality. Some studies show contradicting results of coupling directionality from high frequency to low frequency signal components, in spite of generally considered modulation of a high-frequency amplitude by a low-frequency phase. We have compared two widely used methods to estimate the directionality in cross frequency coupling: conditional mutual information (CMI) and phase slope index (PSI). The latter, applied to infer cross-frequency phase-amplitude directionality from animal intracranial recordings, gives opposite results when comparing to CMI. Both metrics were tested in a numerically simulated example of unidirectionally coupled Rössler systems, which helped to find the explanation of the contradictory results: PSI correctly estimates the lead/lag relationship which, however, is not generally equivalent to causality in the sense of directionality of coupling in nonlinear systems, correctly inferred by using CMI with surrogate data testing.

Dynamic evolution of frontal-temporal network connectivity in temporal lobe epilepsy: A magnetoencephalography study.

Temporal lobe epilepsy (TLE) frequently involves an intricate, extensive epileptic frontal-temporal network. This study aimed to investigate the interactions between temporal and frontal regions and the dynamic patterns of the frontal-temporal network in TLE patients with different disease durations. The magnetoencephalography data of 36 postoperative seizure-free patients with long-term follow-up of at least 1 year, and 21 age- and sex-matched healthy subjects were included in this study. Patients were initially divided into LONG-TERM (n = 18, DURATION >10 years) and SHORT-TERM (n = 18, DURATION ≤10 years) groups based on 10-year disease duration. For reliability, supplementary analyses were conducted with alternative cutoffs, creating three groups: 0 < DURATION ≤7 years (n = 11), 7 < DURATION ≤14 years (n = 11), and DURATION >14 years (n = 14). This study examined the intraregional phase-amplitude coupling (PAC) between theta phase and alpha amplitude across the whole brain. The interregional directed phase transfer entropy (dPTE) between frontal and temporal regions in the alpha and theta bands, and the interregional cross-frequency directionality (CFD) between temporal and frontal regions from the theta phase to the alpha amplitude were further computed and compared among groups. Partial correlation analysis was conducted to investigate correlations between intraregional PAC, interregional dPTE connectivity, interregional CFD, and disease duration. Whole-brain intraregional PAC analyses revealed enhanced theta phase-alpha amplitude coupling within the ipsilateral temporal and frontal regions in TLE patients, and the ipsilateral temporal PAC was positively correlated with disease duration (r = 0.38, p <.05). Interregional dPTE analyses demonstrated a gradual increase in frontal-to-temporal connectivity within the alpha band, while the direction of theta-band connectivity reversed from frontal-to-temporal to temporal-to-frontal as the disease duration increased. Interregional CFD analyses revealed that the inhibitory effect of frontal regions on temporal regions gradually increased with prolonged disease duration (r = -0.36, p <.05). This study clarified the intrinsic reciprocal connectivity between temporal and frontal regions with TLE duration. We propose a dynamically reorganized triple-stage network that transitions from balanced networks to constrained networks and further develops into imbalanced networks as the disease duration increases.

Global Run-On sequencing to measure nascent transcription in Saccharomyces cerevisiae.

Global Run-On sequencing is a reliable and widely used approach for monitoring nascent transcription on a genomewide scale. The assay has been successfully used for studying global transcription in humans, plants, worms, flies, and fission yeast. Here we describe a GRO-seq protocol for studying transcription in budding yeast, Saccharomyces cerevisiae. Briefly, the technique involves permeabilization of actively growing yeast cells, allowing transcription to proceed in permeabilized cells in the presence of brominated UTP, affinity purification of bromo-UMP incorporated nascent transcripts followed by cDNA library construction, deep sequencing, and mapping against the reference genome. The approach maps the position of transcriptionally active RNA polymerase on a genomewide basis. In addition to identifying the complete set of transcriptionally active genes in a cell under a given set of conditions, the method can be used to determine elongation rate, termination defect and promoter directionality at the genomewide level. The approach is especially useful in identifying short-lived unstable transcripts that are rapidly degraded even before they leave the nucleus.

Effects of immediate loading directionality on the mechanical sensing protein PIEZO1 expression and early-stage healing process of peri-implant bone.

BACKGROUND: The reduced treatment time of dental implants with immediate loading protocol is an appealing solution for dentists and patients. However, there remains a significant risk of early peri-implant bone response following the placement of immediately loaded implants, and limited information is available regarding loading directions and the associated in vivo characteristics of peri-implant bone during the early stages. This study aimed to investigate the effects of immediate loading directionality on the expression of mechanical sensing protein PIEZO1 and the healing process of peri-implant bone in the early stage. METHODS: Thirty-two implants were inserted into the goat iliac crest models with 10 N static lateral immediate loading applied, followed by histological, histomorphological, immunohistochemical, X-ray microscopy and energy dispersive X-ray spectroscopy evaluations conducted after 10 days. RESULTS: From evaluations at the cellular, tissue, and organ levels, it was observed that the expression of mechanical sensing protein PIEZO1 in peri-implant bone was significantly higher in the compressive side compared to the tensile side. This finding coincided with trends observed in interfacial bone extracellular matrix (ECM) contact percentage, bone mass, and new bone formation. CONCLUSIONS: This study provides a novel insight into the immediate loading directionality as a potential influence factor for dental implant treatments by demonstrating differential effects on the mechanical sensing protein PIEZO1 expression and related early-stage healing processes of peri-implant bone. Immediate loading directions serve as potential therapeutic influence factors for peri-implant bone during its early healing stage.

Efficient conversion of hemoglobin to a non-vasoactive oxygen carrier by site-specific cross-linking with azido acyl methyl phosphates followed by bio-orthogonal CuAAC with a bis-alkyne.

While cross-linked hemoglobin tetramers are functional acellular oxygen carriers, their ability to scavenge endogenous nitric oxide (NO) by endothelial pore penetration results in adverse cardiovascular effects. Animal studies established that cross-linked human hemoglobins, chemically joined into a double protein, avoid NO scavenging, presumably due to their larger size preventing penetration into endothelial regions that produce NO. In the present report, we utilize azide-containing acyl phosphate reagents to form cross-linked hemoglobins then bio-orthogonally click-couple them with a bis-alkyne (CuAAC). The production of these larger oxygen-carrying hemoglobin conjugates is obtained in high yields through subunit-specific cross-linking between each ßLys82 ε-amino group. The methyl phosphate leaving groups provide electrostatically induced ß-subunit site-selectivity, producing azido-cross-linked hemoglobin that undergoes highly efficient CuAAC compared with previous cross-linkers. The acyl phosphates also efficiently cross-link both T-state and R-state hemoglobin. The resulting bis- and tris-tetrameric hemoglobin conjugates exhibit oxygen affinity and cooperativity that are comparable to those of the native protein. The hemoglobin derivatives from the process we describe can function as sources of oxygen in biomedical applications, such as in ex-vivo donor organ perfusion.

Intergenerational transmission of comorbid internalizing and externalizing psychopathology at age 11: Evidence from an adoption design for general transmission of comorbidity rather than homotypic transmission.

Psychopathology is intergenerationally transmitted through both genetic and environmental mechanisms via heterotypic (cross-domain), homotypic (domain-specific), and general (e.g., "p-factor") pathways. The current study leveraged an adopted-at-birth design, the Early Growth and Development Study (57% male; 55.6% White, 19.3% Multiracial, 13% Black/African American, 10.9% Hispanic/Latine) to explore the relative influence of these pathways via associations between adoptive caregiver psychopathology (indexing potential environmental transmission) and birth parent psychopathology (indexing genetic transmission) with adolescent internalizing and externalizing symptoms. We included composite measures of adoptive and birth parent internalizing, externalizing, and substance use domains, and a general "p-factor." Age 11 adolescent internalizing and externalizing symptom scores were the average of adoptive parent reports on the Child Behavior Checklist (n = 407). Examining domains independently without addressing comorbidity can lead to incorrect interpretations of transmission mode. Therefore, we also examined symptom severity (like the "p-factor") and an orthogonal symptom directionality score to more cleanly disentangle transmission modes. The pattern of correlations was consistent with mostly general transmission in families with youth showing comorbid internalizing and externalizing symptoms, rather than homotypic transmission. Findings more strongly supported potential environmental or evocative mechanisms of intergenerational transmission than genetic transmission mechanisms (though see limitations). Parent-specific effects are discussed.

A specialized spinal circuit for command amplification and directionality during escape behavior.

In vertebrates, action selection often involves higher cognition entailing an evaluative process. However, urgent tasks, such as defensive escape, require an immediate implementation of the directionality of escape trajectory, necessitating local circuits. Here we reveal a specialized spinal circuit for the execution of escape direction in adult zebrafish. A central component of this circuit is a unique class of segmentally repeating cholinergic V2a interneurons expressing the transcription factor Chx10. These interneurons amplify brainstem-initiated escape commands and rapidly deliver the excitation via a feedforward circuit to all fast motor neurons and commissural interneurons to direct the escape maneuver. The information transfer within this circuit relies on fast and reliable axo-axonic synaptic connections, bypassing soma and dendrites. Unilateral ablation of cholinergic V2a interneurons eliminated escape command propagation. Thus, in vertebrates, local spinal circuits can implement directionality of urgent motor actions vital for survival.

Bidirectional Within-Family Effects of Restrictive Mediation Practices and Adolescents' Problematic Social Media Use.

Much remains unknown about whether restrictive mediation is an effective parenting strategy to prevent or reduce problematic social media use among adolescents. Therefore, this study examined bidirectional within-family effects between two restrictive mediation practices (rule-setting and reactive restrictions) and problematic social media use using random-intercept cross-lagged panel modeling. Three-wave survey data collected among Dutch adolescents (T1: N = 1928, Mage = 13.31 years, SD = 0.91, 43.3% girl) with a 1 year-interval were used. Results showed that within-family changes in problematic social media use symptoms predicted subsequent within-family changes in perceived parental restrictive mediation. More specifically, an increase in symptoms predicted a decrease in rule-setting and an increase in reactive restrictions 1 year later. Within-family changes in perceived parental restrictive mediation practices did not predict within-family changes in problematic social media use symptoms, suggesting that the relation is unidirectional. However, concluding that limiting adolescents' Internet use is ineffective to prevent problematic social media use would be premature. Future research should investigate whether it may be an effective parenting strategy for a certain subgroup of adolescents or under certain circumstances.

Temporal Directionality Between Symptoms During Treatment of Depressed Inpatients: A Dynamic Time Warp Network Analysis.

BACKGROUND: Depression is generally perceived from the perspective of the common-cause disease model. However, the network perspective assumes mutual influence of individual symptoms and stresses the importance of investigating symptom dynamics. Gaining a better understanding of symptom dynamics within individuals might contribute to more effective treatments. METHODS: Current exploratory longitudinal research studied the associations and directionality between 43 symptoms from the generic questionnaire Symptom Questionnaire-48 (SQ-48) using dynamic time warp (DTW) analyses, in which trajectories with similar time-dependent patterns can be identified. Data from individuals were analysed first, yielding distance matrices for all symptom trajectories, after which the data were aggregated. RESULTS: The 148 included patients were all admitted for the treatment of their clinical depression. Undirected DTW analyses of three patients with longer time series but otherwise randomly chosen showed large variability among individuals. Group-level undirected DTW analyses showed numerous significant edges between symptoms, largely clustering symptoms according to the eight pre-existing subscales of the SQ-48. Group-level directed DTW analyses showed five symptoms with significant outstrength: 'hopeless', 'restless', 'down/depressed', 'feeling tense' and 'no enjoyment', meaning that change in these key symptoms preceded change in other symptoms. LIMITATIONS: The 43 included symptoms of the SQ-48 primarily focus on internalizing problems in severely depressed inpatients, potentially limiting generalizability. CONCLUSIONS: DTW networks provided us with five key symptoms based on the dynamics of symptom scores. Future studies could explore whether process-based therapy targeted at symptoms with high outstrength might result in more effectivity as part of personalized treatment.

Deep learning-enabled natural language processing to identify directional pharmacokinetic drug-drug interactions.

BACKGROUND: During drug development, it is essential to gather information about the change of clinical exposure of a drug (object) due to the pharmacokinetic (PK) drug-drug interactions (DDIs) with another drug (precipitant). While many natural language processing (NLP) methods for DDI have been published, most were designed to evaluate if (and what kind of) DDI relationships exist in the text, without identifying the direction of DDI (object vs. precipitant drug). Here we present a method for the automatic identification of the directionality of a PK DDI from literature or drug labels. METHODS: We reannotated the Text Analysis Conference (TAC) DDI track 2019 corpus for identifying the direction of a PK DDI and evaluated the performance of a fine-tuned BioBERT model on this task by following the training and validation steps prespecified by TAC. RESULTS: This initial attempt showed the model achieved an F-score of 0.82 in identifying sentences as containing PK DDI and an F-score of 0.97 in identifying object versus precipitant drugs in those sentences. DISCUSSION AND CONCLUSION: Despite a growing list of NLP methods for DDI extraction, most of them use a common set of corpora to perform general purpose tasks (e.g., classifying a sentence into one of several fixed DDI categories). There is a lack of coordination between the drug development and biomedical informatics method development community to develop corpora and methods to perform specific tasks (e.g., extract clinical exposure changes due to PK DDI). We hope that our effort can encourage such a coordination so that more "fit for purpose" NLP methods could be developed and used to facilitate the drug development process.

Identity Formation and General and Cancer-specific Functioning in Adolescent and Emerging Adult Survivors of Childhood Cancer: A Longitudinal Study into Directionality of Effects.

BACKGROUND: Adolescent and emerging adult survivors of childhood cancer generally adjust well psychologically similar to their peers. Nevertheless, some survivors are at greater risk for developing psychological and physical difficulties. To shed light on the psychosocial functioning of adolescent and emerging adult survivors of childhood cancer, personal identity formation and its interplay with general and cancer-specific functioning need to be investigated. PURPOSE: To examine the longitudinal associations linking identity formation to general and cancer-specific functioning in adolescent and emerging adult childhood cancer survivors using three-wave data over a 2-year period. METHODS: Dutch-speaking survivors (at baseline: n = 125; 53% female; age range: 14-25 years) treated at the pediatric oncology department of the University Hospitals Leuven (Belgium), completed self-report questionnaires at three annual timepoints. Directionality of effects and correlated changes were examined using cross-lagged structural equation modeling. RESULTS: Regarding general functioning, bidirectional effects occurred. Life satisfaction positively predicted identity synthesis and both life satisfaction and good physical functioning negatively predicted identity confusion over time. Identity synthesis, in turn, positively predicted life satisfaction and identity confusion negatively predicted good physical functioning over time. Regarding cancer-specific functioning, mainly unidirectional effects occurred. Post-traumatic stress symptoms negatively predicted identity synthesis and positively predicted identity confusion over time, whereas the reverse pattern of associations was found for benefit finding. Several correlated changes were found linking identity formation and psychosocial functioning as well. CONCLUSIONS: The present study uncovered clinically meaningful pathways linking identity formation to psychosocial functioning over time in adolescents and emerging adults who survived childhood cancer.

To shed light on the psychosocial functioning of adolescent and emerging adult survivors of childhood cancer, personal identity formation and its longitudinal interplay with general and cancer-specific functioning need to be investigated. Dutch-speaking survivors treated at the pediatric oncology department of the University Hospitals Leuven (Belgium), completed self-report questionnaires at three annual timepoints, resulting in three-wave data over a 2-year period. Regarding identity formation and general functioning, bidirectional effects occurred. Life satisfaction positively predicted identity synthesis and both life satisfaction and good physical functioning negatively predicted identity confusion over time. Identity synthesis, in turn, positively predicted life satisfaction and identity confusion negatively predicted good physical functioning over time. Regarding identity formation and cancer-specific functioning, mainly unidirectional effects occurred. Post-traumatic stress symptoms negatively predicted identity synthesis and positively predicted identity confusion over time, whereas the reverse pattern of associations was found for benefit finding. The present study uncovered meaningful pathways linking identity formation to psychosocial functioning over time in adolescents and emerging adults who survived childhood cancer. These longitudinal findings may provide important guidance for clinical practice, given that identity formation in today's western society has become particularly challenging.

Investigating the cognitive processing of tools: Effects of dangerousness and directionality on attentional biases.

Research has found a tool-based head-superiority effect (responses are faster to target stimuli that appear at the head than at the handle position). However, these studies did not consider the effects of directionality and dangerousness. This study investigated how directionality and dangerousness influence the head-superiority effect. Subjects were required to respond to the target location in all experiments. Experiment 1 manipulated the directionality, dangerousness and target location. Experiment 2 matched the sharpness of the tool tip in the directed conditions. Experiment 3 shortened the presentation time of the cue stimuli from 800 ms to 200 ms. Experiment 4 selected four tools with different functions to rule out an alternative explanation caused by functional repetition. The results indicate that a head-superiority effect emerges in the directed condition, and that dangerousness modulates the magnitude of this effect during the 800 ms condition. However, the modulation effect of dangerousness diminishes during the 200 ms condition.

Anger dysregulation and non-suicidal self-injury during adolescence: A test of directionality.

Non-suicidal self-injury (NSSI) has been tied to several forms of emotional and behavioral dysregulation in adolescence, with less attention paid to regulation of anger. Most assume that anger dysregulation leads to engagement in NSSI, rather than the reverse. However, it is plausible that NSSI compromises adolescents' abilities to regulate their emotions, including anger, because it may reduce the development of alternative regulatory strategies and intensify negative emotions by reducing tolerance of distress. Using three waves of data from a sample of adolescents in 17 Swedish schools (n = 1,304 M age = 13.68, SD age = .67; 89% of Swedish origin; 58% girls), we examined the directionality of ties between NSSI and three forms of anger dysregulation: dysregulated expressions of anger, anger suppression, and low anger reflection. We also looked for differences in magnitude of paths and gender differences. Random-intercept cross-lagged panel models showed that NSSI predicted changes in all forms of anger dysregulation but found no support for the opposite direction. Gender differences were not evident. Results challenge directionality assumptions and support suggestions that adolescents' anger regulation degrades when they self-injure.

Simultaneous learning of directional and non-directional stimulus relations in baboons (Papio papio).

While humans exposed to a sequential stimulus pairing A-B are commonly assumed to form a bidirectional mental relation between A and B, evidence that non-human animals can do so is limited. Careful examination of the animal literature suggests possible improvements in the test procedures used to probe such effects, notably measuring transfer effects on the learning of B-A pairings, rather than direct recall of A upon cuing with B. We developed such an experimental design and tested 20 Guinea baboons (Papio papio). Two pairings of visual shapes were trained (A1-B1, A2-B2) and testing was conducted in a reversed order, either with conserved pairings (B1-A1, B2-A2) or broken ones (B1-A2, B2-A1). We found baboons' immediate test performance to be above chance level for conserved pairings and below chance level for broken ones. Moreover, baboons needed less trials to learn conserved pairings compared to broken ones. These effects were apparent for both pairings on average, and separately for the best learned pairing. Baboons' responding on B-A trials was thus influenced by their previous A-B training. Performance level at the onset of testing, however, suggests that baboons did not respond in full accordance with the hypothesis of bidirectionality. To account for these data, we suggest that two competing types of relations were concomitantly encoded: a directional relation between A and B, which retains the sequential order experienced, and a non-directional relation, which retains only the co-occurrence of events, not their temporal order.