Common and unique neurophysiological signatures for the stopping and revising of actions reveal the temporal dynamics of inhibitory control.

Inhibitory control is a crucial cognitive-control ability for behavioral flexibility that has been extensively investigated through action-stopping tasks. Multiple neurophysiological features have been proposed to represent 'signatures' of inhibitory control during action-stopping, though the processes signified by these signatures are still controversially discussed. The present study aimed to disentangle these processes by comparing simple stopping situations with those in which additional action revisions were needed. Three experiments in female and male humans were performed to characterize the neurophysiological dynamics involved in action-stopping and - changing, with hypotheses derived from recently developed two-stage 'pause-then-cancel' models of inhibitory control. Both stopping and revising an action triggered an early broad 'pause'-process, marked by frontal EEG ß-bursts and non-selective suppression of corticospinal excitability. However, partial-EMG responses showed that motor activity was only partially inhibited by this 'pause', and that this activity can be further modulated during action-revision. In line with two-stage models of inhibitory control, subsequent frontocentral EEG activity after this initial 'pause' selectively scaled depending on the required action revisions, with more activity observed for more complex revisions. This demonstrates the presence of a selective, effector-specific 'retune' phase as the second process involved in action-stopping and -revision. Together, these findings show that inhibitory control is implemented over an extended period of time and in at least two phases. We are further able to align the most commonly proposed neurophysiological signatures to these phases and show that they are differentially modulated by the complexity of action-revision. Significance Statement: Inhibitory control is one of the most important control processes by which humans can regulate their behavior. Multiple neurophysiological signatures have been proposed to reflect inhibitory control. However, these play out on different time scales and appear to reflect different aspects of cognitive control, which are controversially debated.Recent two-stage models of inhibitory control have proposed that two phases implement the revisions of actions: 'pause' and 'retune'. Here, we provide the first empirical evidence for this proposition: Action revisions engendered a common initial low-latency 'pause', during which motor activity is broadly suppressed. Later activity, however, distinguishes between simple stopping of actions and more complex action revisions. These findings provide novel insights into the sequential dynamics of human action control.

Pyridopyrimidinones as a new chemotype of calcium dependent protein kinase 1 (CDPK1) inhibitors for Cryptosporidium.

The protozoan protein kinase calcium-dependent protein kinase 1 (CDPK1) has emerged as a potential therapeutic target for the treatment of cryptosporidiosis. A focused screen of known kinase inhibitors identified a pyridopyrimidinone as a new chemotype of Cryptosporidium parvum (Cp) CDPK1 inhibitors. Structural comparison of CpCDPK1 to two representative human kinases, RIPK2 and Src, revealed differences in the positioning of the αC-helix that was used in the design of a potent pyridopyrimidinone-based CpCDPK1 inhibitor 7 (a.k.a. UH15-16, IC50 = 10 nM), which blocked the growth of three C. parvum strains (EC50 = 12-40 nM) as well as C. hominis (EC50 = 85 nM) in HCT-8 host cells. Pharmacokinetic and tissue distribution analyses indicated that 7 had low systemic exposure after oral administration, but high gastrointestinal concentration, as well as good Caco-2 cell permeability. Finally, 7 demonstrated partial efficacy in an IL-12 knock-out mouse model of acute cryptosporidiosis.

Randomised study of the effects of sense of entitlement and conflict of interest contrarianism on researcher decision-making to work with the alcohol industry.

BACKGROUND: It is well established that the tobacco industry used research funding as a deliberate tactic to subvert science. There has been little wider attention to how researchers think about accepting industry funding. We developed, then tested, hypotheses about two psychological constructs, namely, entitlement and conflict of interest contrarianism (CoI-C) among alcohol researchers who had previously received industry funding. METHODS: A mixed-methods pilot study involved construct and instrument development, followed by an online survey and nested 3-arm randomised trial. We randomly allocated alcohol industry funding recipients to one of three conditions. In two experimental conditions we asked participants questions to remind them (and thus increase the salience) of their sense of entitlement or CoI-C. We compared these groups with a control group who did not receive any reminder. The outcome was a composite measure of openness to working with the alcohol industry. RESULTS: 133 researchers were randomised of whom 79 completed the experiment. The posterior distribution over effect estimates revealed that there was a 94.8% probability that reminding researchers of their CoI-C led them to self-report being more receptive to industry funding, whereas the probability was 68.1% that reminding them of their sense of entitlement did so. Biomedical researchers reported being more open to working with industry than did psychosocial researchers. CONCLUSION: Holding contrarian views on conflict of interest could make researchers more open to working with industry. This study shows how it is possible to study researcher decision-making using quantitative experimental methods.

Strategies, processes, outcomes, and costs of implementing experience sampling-based monitoring in routine mental health care in four European countries: study protocol for the IMMERSE effectiveness-implementation study.

BACKGROUND: Recent years have seen a growing interest in the use of digital tools for delivering person-centred mental health care. Experience Sampling Methodology (ESM), a structured diary technique for capturing moment-to-moment variation in experience and behaviour in service users' daily life, reflects a particularly promising avenue for implementing a person-centred approach. While there is evidence on the effectiveness of ESM-based monitoring, uptake in routine mental health care remains limited. The overarching aim of this hybrid effectiveness-implementation study is to investigate, in detail, reach, effectiveness, adoption, implementation, and maintenance as well as contextual factors, processes, and costs of implementing ESM-based monitoring, reporting, and feedback into routine mental health care in four European countries (i.e., Belgium, Germany, Scotland, Slovakia). METHODS: In this hybrid effectiveness-implementation study, a parallel-group, assessor-blind, multi-centre cluster randomized controlled trial (cRCT) will be conducted, combined with a process and economic evaluation. In the cRCT, 24 clinical units (as the cluster and unit of randomization) at eight sites in four European countries will be randomly allocated using an unbalanced 2:1 ratio to one of two conditions: (a) the experimental condition, in which participants receive a Digital Mobile Mental Health intervention (DMMH) and other implementation strategies in addition to treatment as usual (TAU) or (b) the control condition, in which service users are provided with TAU. Outcome data in service users and clinicians will be collected at four time points: at baseline (t0), 2-month post-baseline (t1), 6-month post-baseline (t2), and 12-month post-baseline (t3). The primary outcome will be patient-reported service engagement assessed with the service attachment questionnaire at 2-month post-baseline. The process and economic evaluation will provide in-depth insights into in-vivo context-mechanism-outcome configurations and economic costs of the DMMH and other implementation strategies in routine care, respectively. DISCUSSION: If this trial provides evidence on reach, effectiveness, adoption, implementation and maintenance of implementing ESM-based monitoring, reporting, and feedback, it will form the basis for establishing its public health impact and has significant potential to bridge the research-to-practice gap and contribute to swifter ecological translation of digital innovations to real-world delivery in routine mental health care. TRIAL REGISTRATION: ISRCTN15109760 (ISRCTN registry, date: 03/08/2022).

In Support of Multidimensional Frailty: A Structural Equation Model from the Canadian Positive Brain Health Now Cohort.

The objective of this study was to estimate the structure and relationships between four h ypothesized frailty dimensions (physical, emotional, cognitive, and social) and the extent to which personal and HIV-related factors and comorbidity associate with these frailty dimensions. This is a secondary analysis of an existing dataset arising from Positive Brain Health Now study (n = 856) in people aging with HIV (mean age: 52.3 ± 8.1 years). Structural equation modeling (SEM) models were applied to two cross-sections of the data: one at study entry and one at second visit, 9-month apart. Multidimensional frailty was modeled based on the combined Wilson-Cleary and International Classification of Functioning, Disability and Health framework. Four dimensions were operationalized with patient-reported and self-report measures from standardized questionnaires. The SEM model from the first visit was replicated using data from the second visit, testing measurement invariance. The proposed model showed acceptable fit at both visits (including no violation of measurement invariance). The final model for the first visit showed that sex, body mass index, HIV diagnosis pre-1997, current or nadir CD4 counts, and comorbidity did not associate with any frailty dimension; however, age (ß range: 0.12-0.25), symptoms (ß range: -0.35 to -0.58), and measured cognition (ß range: 0.10-0.24) directly associated with all frailty dimensions. The model remained stable across the two visits. This study contributes evidence for operationalizing multidimensional frailty. Evidence-based interventions are available for many of the measures considered here, offering opportunities to improve the lives of people with frailty in the context of HIV.

Effect of N Atom Substitution on Electronic Resonances: A 2D Photoelectron Spectroscopic and Computational Study of Anthracene, Acridine, and Phenazine Anions.

The accommodation of an excess electron by polycyclic aromatic hydrocarbons (PAHs) has important chemical and technological implications ranging from molecular electronics to charge balance in interstellar molecular clouds. Here, we use two-dimensional photoelectron spectroscopy and equation-of-motion coupled-cluster calculations of the radical anions of acridine (C13H9N-) and phenazine (C12H8N2-) and compare our results for these species to those for the anthracene anion (C14H10-). The calculations predict the observed resonances and additionally find low-energy two-particle-one-hole states, which are not immediately apparent in the spectra, and offer a slightly revised interpretation of the resonances in anthracene. The study of acridine and phenazine allows us to understand how N atom substitution affects electron accommodation. While the electron affinity associated with the ground state anion undergoes a sizable increase with the successive substitution of N atoms, the two lowest energy excited anion states are not affected significantly by the substitution. The net result is that there is an increase in the energy gap between the two lowest energy resonances and the bound ground electronic state of the radical anion from anthracene to acridine to phenazine. Based on an energy gap law for the rate of internal conversion, this increased gap makes ground state formation progressively less likely, as evidenced by the photoelectron spectra.

A Guide to Measuring Heart and Respiratory Rates Based on Off-the-Shelf Photoplethysmographic Hardware and Open-Source Software.

The remote monitoring of vital signs via wearable devices holds significant potential for alleviating the strain on hospital resources and elder-care facilities. Among the various techniques available, photoplethysmography stands out as particularly promising for assessing vital signs such as heart rate, respiratory rate, oxygen saturation, and blood pressure. Despite the efficacy of this method, many commercially available wearables, bearing Conformité Européenne marks and the approval of the Food and Drug Administration, are often integrated within proprietary, closed data ecosystems and are very expensive. In an effort to democratize access to affordable wearable devices, our research endeavored to develop an open-source photoplethysmographic sensor utilizing off-the-shelf hardware and open-source software components. The primary aim of this investigation was to ascertain whether the combination of off-the-shelf hardware components and open-source software yielded vital-sign measurements (specifically heart rate and respiratory rate) comparable to those obtained from more expensive, commercially endorsed medical devices. Conducted as a prospective, single-center study, the research involved the assessment of fifteen participants for three minutes in four distinct positions, supine, seated, standing, and walking in place. The sensor consisted of four PulseSensors measuring photoplethysmographic signals with green light in reflection mode. Subsequent signal processing utilized various open-source Python packages. The heart rate assessment involved the comparison of three distinct methodologies, while the respiratory rate analysis entailed the evaluation of fifteen different algorithmic combinations. For one-minute average heart rates' determination, the Neurokit process pipeline achieved the best results in a seated position with a Spearman's coefficient of 0.9 and a mean difference of 0.59 BPM. For the respiratory rate, the combined utilization of Neurokit and Charlton algorithms yielded the most favorable outcomes with a Spearman's coefficient of 0.82 and a mean difference of 1.90 BrPM. This research found that off-the-shelf components are able to produce comparable results for heart and respiratory rates to those of commercial and approved medical wearables.

Evidence for a role of synchrony but not common fate in the perception of biological group movements.

Extensive research has shown that observers are able to efficiently extract summary information from groups of people. However, little is known about the cues that determine whether multiple people are represented as a social group or as independent individuals. Initial research on this topic has primarily focused on the role of static cues. Here, we instead investigate the role of dynamic cues. In two experiments with male and female human participants, we use EEG frequency tagging to investigate the influence of two fundamental Gestalt principles - synchrony and common fate - on the grouping of biological movements. In Experiment 1, we find that brain responses coupled to four point-light figures walking together are enhanced when they move in sync vs. out of sync, but only when they are presented upright. In contrast, we found no effect of movement direction (i.e., common fate). In Experiment 2, we rule out that synchrony takes precedence over common fate by replicating the null effect of movement direction while keeping synchrony constant. These results suggest that synchrony plays an important role in the processing of biological group movements. In contrast, the role of common fate is less clear and will require further research.

Attentional set and explicit expectations of perceptual load determine flanker interference.

Task-irrelevant stimuli often capture our attention despite our best efforts to ignore them. It has been noted that tasks involving perceptually complex displays can lead to reduced interference from distractors. The mechanism behind this effect is debated, with some accounts emphasizing the "perceptual load" of the stimuli themselves and others emphasizing the role of proactive control. Here, in three experiments, we investigated the roles of perceptual load, proactive control, and reward motivation in determining distractor interference. Participants performed a visual search task of high, low, or intermediate load, with flanking task-irrelevant distractors. Each trial was preceded by a cue indicating the level of perceptual load (Experiments 1-3) as well as the potential reward that could be earned (Experiments 2 and 3). In all three experiments, the attentional set induced by the preceding trial and cued proactive expectation of perceptual load interacted to determine flanker interference, which was significant for all trial types except trials cued as high load which were also preceded by high load. These effects were not modulated by reward motivation, although in the final experiment reward did significantly improve performance overall. Thus, successful distractor exclusion does not depend upon motivation or load per se but does require an expectation of high load. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Top-down biological motion perception does not differ between adults scoring high versus low on autism traits.

The perception of biological motion is an important social cognitive ability. Models of biological motion perception recognize two processes that contribute to the perception of biological motion: a bottom-up process that binds optic-flow patterns into a coherent percept of biological motion and a top-down process that binds sequences of body-posture 'snapshots' over time into a fluent percept of biological motion. The vast majority of studies on autism and biological motion perception have used point-light figure stimuli, which elicit biological motion perception predominantly via bottom-up processes. Here, we investigated whether autism is associated with deviances in the top-down processing of biological motion. For this, we tested a sample of adults scoring low vs high on autism traits on a recently validated EEG paradigm in which apparent biological motion is combined with frequency tagging (Cracco et al., 2022) to dissociate between two percepts: 1) the representation of individual body postures, and 2) their temporal integration into movements. In contrast to our hypothesis, we found no evidence for a diminished temporal body posture integration in the high-scoring group. We did, however, find a group difference that suggests that adults scoring high on autism traits have a visual processing style that focuses more on a single percept (i.e. either body postures or movements, contingent on saliency) compared to adults scoring low on autism traits who instead seemed to represent the two percepts included in the paradigm in a more balanced manner. Although unexpected, this finding aligns well with the autism literature on perceptual stability.

The LEADING Guideline: Reporting Standards for Expert Panel, Best-Estimate Diagnosis, and Longitudinal Expert All Data (LEAD) Studies.

Accurate assessments of symptoms and diagnoses are essential for health research and clinical practice but face many challenges. The absence of a single error-free measure is currently addressed by assessment methods involving experts reviewing several sources of information to achieve a more accurate or best-estimate assessment. Three bodies of work spanning medicine, psychiatry, and psychology propose similar assessment methods: The Expert Panel, the Best-Estimate Diagnosis, and the Longitudinal Expert All Data (LEAD). However, the quality of such best-estimate assessments is typically very difficult to evaluate due to poor reporting of the assessment methods and when it is reported, the reporting quality varies substantially. Here we tackle this gap by developing reporting guidelines for such studies, using a four-stage approach: 1) drafting reporting standards accompanied by rationales and empirical evidence, which were further developed with a patient organization for depression, 2) incorporating expert feedback through a two-round Delphi procedure, 3) refining the guideline based on an expert consensus meeting, and 4) testing the guideline by i) having two researchers test it and ii) using it to examine the extent previously published articles report the standards. The last step also demonstrates the need for the guideline: 18 to 58% (Mean = 33%) of the standards were not reported across fifteen randomly selected studies. The LEADING guideline comprises 20 reporting standards related to four groups: The Longitudinal design; the Appropriate data; the Evaluation - experts, materials, and procedures; and the Validity group. We hope that the LEADING guideline will be useful in assisting researchers in planning, reporting, and evaluating research aiming to achieve best-estimate assessments.

Photoelectron spectroscopy of the deprotonated tryptophan anion: the contribution of deprotomers to its photodetachment channels.

Photoelectron spectroscopy and electronic structure calculations are used to investigate the electronic structure of the deprotonated anionic form of the aromatic amino acid tryptophan, and its chromophore, indole. The photoelectron spectra of tryptophan, recorded at different wavelengths across the UV, consist of two direct detachment channels and thermionic emission, whereas the hν = 4.66 eV spectrum of indole consists of two direct detachment features. Electronic structure calculations indicate that two deprotomers of tryptophan are present in the ion beam; deprotonation of the carboxylic acid group (Trp(I)-) or the N atom on the indole ring (Trp(II)-). Strong similarities are observed between the direct detachment channels in the photoelectron spectra of tryptophan and indole, which in conjunction with electronic structure calculations, indicate that electron loss from Trp(II)- dominates this portion of the spectra. However, there is some evidence that direct detachment of Trp(I)- is also observed. Thermionic emission is determined to predominantly arise from the decarboxylation of Trp(I)-, mediated by the ππ* excited state near λ = 300 nm, which results in an anionic fragment with a negative electron affinity that readily autodetaches.

Biophysical Modeling of Frontocentral ERP Generation Links Circuit-Level Mechanisms of Action-Stopping to a Behavioral Race Model.

Human frontocentral event-related potentials (FC-ERPs) are ubiquitous neural correlates of cognition and control, but their generating multiscale mechanisms remain mostly unknown. We used the Human Neocortical Neurosolver's biophysical model of a canonical neocortical circuit under exogenous thalamic and cortical drive to simulate the cell and circuit mechanisms underpinning the P2, N2, and P3 features of the FC-ERP observed after Stop-Signals in the Stop-Signal task (SST; N = 234 humans, 137 female). We demonstrate that a sequence of simulated external thalamocortical and corticocortical drives can produce the FC-ERP, similar to what has been shown for primary sensory cortices. We used this model of the FC-ERP to examine likely circuit-mechanisms underlying FC-ERP features that distinguish between successful and failed action-stopping. We also tested their adherence to the predictions of the horse-race model of the SST, with specific hypotheses motivated by theoretical links between the P3 and Stop process. These simulations revealed that a difference in P3 onset between successful and failed Stops is most likely due to a later arrival of thalamocortical drive in failed Stops, rather than, for example, a difference in the effective strength of the input. In contrast, the same model predicted that early thalamocortical drives underpinning the P2 and N2 differed in both strength and timing across stopping accuracy conditions. Overall, this model generates novel testable predictions of the thalamocortical dynamics underlying FC-ERP generation during action-stopping. Moreover, it provides a detailed cellular and circuit-level interpretation that supports links between these macroscale signatures and predictions of the behavioral race model.

Infrared Thermographic Imaging as a Tool to Assess Inflammatory and Ischemic Response With Rubber Ring Tail Docking in Suffolk and Hampshire Lambs (Ovis Aries).

Tail docking in lambs is associated with an acute inflammatory and ischemic response correlated to pain. The present study sought to quantify these responses by utilizing infrared thermographic imaging (IRT) after tail docking lambs. Images were taken for each lamb before tail docking (T0), within a minute after the band was placed (T1), at 5 minutes (T5), and 15 minutes (T15) respectively and analyzed for pixel intensity in areas on the lamb's rump, tail base, and two locations below the docking band. The pixel intensity at 2 cm below the docking band did significantly decrease (p < 0.05) over the course of the study at 15 minutes, indicating the presence of ischemia, and a significant decrease in the pixel intensity of the rump and tail base was observed. The closest location to the rubber ring showed the presence of inflammation as an indirect indicator of acute pain . IRT is a valuable tool in the detection of inflammation and ischemic areas and has the potential to be used to quantify and measure the experience of inflammation and pain after tail docking.

The human subthalamic nucleus transiently inhibits active attentional processes.

The subthalamic nucleus (STN) of the basal ganglia is key to the inhibitory control of movement. Consequently, it is a primary target for the neurosurgical treatment of movement disorders like Parkinson's Disease, where modulating the STN via deep-brain stimulation (DBS) can release excess inhibition of thalamo-cortical motor circuits. However, the STN is also anatomically connected to other thalamo-cortical circuits, including those underlying cognitive processes like attention. Notably, STN-DBS can also affect these processes. This suggests that the STN may also contribute to the inhibition of non-motor activity, and that STN-DBS may cause changes to this inhibition. We here tested this hypothesis in humans. We used a novel, wireless outpatient method to record intracranial local field potentials (LFP) from STN DBS implants during a visual attention task (Experiment 1, N=12). These outpatient measurements allowed the simultaneous recording of high-density EEG, which we used to derive the steady-state visual evoked potential (SSVEP), a well-established neural index of visual attentional engagement. By relating STN activity to this neural marker of attention (instead of overt behavior), we avoided possible confounds resulting from STN's motor role. We aimed to test whether the STN contributes to the momentary inhibition of the SSVEP caused by unexpected, distracting sounds. Furthermore, we causally tested this association in a second experiment, where we modulated STN via DBS across two sessions of the task, spaced at least one week apart (N=21, no sample overlap with Experiment 1). The LFP recordings in Experiment 1 showed that reductions of the SSVEP after distracting sounds were preceded by sound-related γ-frequency (>60Hz) activity in the STN. Trial-to-trial modeling further showed that this STN activity statistically mediated the sounds' suppressive effect on the SSVEP. In Experiment 2, modulating STN activity via DBS significantly reduced these sound-related SSVEP reductions. This provides causal evidence for the role of the STN in the surprise-related inhibition of attention. These findings suggest that the human STN contributes to the inhibition of attention, a non-motor process. This supports a domain-general view of the inhibitory role of the STN. Furthermore, these findings also suggest a potential mechanism underlying some of the known cognitive side-effects of STN-DBS treatment, especially on attentional processes. Finally, our newly-established outpatient LFP recording technique facilitates the testing of the role of subcortical nuclei in complex cognitive tasks, alongside recordings from the rest of the brain, and in much shorter time than perisurgical recordings.

Tomosyns attenuate SNARE assembly and synaptic depression by binding to VAMP2-containing template complexes.

Tomosyns are widely thought to attenuate membrane fusion by competing with synaptobrevin-2/VAMP2 for SNARE-complex assembly. Here, we present evidence against this scenario. In a novel mouse model, tomosyn-1/2 deficiency lowered the fusion barrier and enhanced the probability that synaptic vesicles fuse, resulting in stronger synapses with faster depression and slower recovery. While wild-type tomosyn-1m rescued these phenotypes, substitution of its SNARE motif with that of synaptobrevin-2/VAMP2 did not. Single-molecule force measurements indeed revealed that tomosyn's SNARE motif cannot substitute synaptobrevin-2/VAMP2 to form template complexes with Munc18-1 and syntaxin-1, an essential intermediate for SNARE assembly. Instead, tomosyns extensively bind synaptobrevin-2/VAMP2-containing template complexes and prevent SNAP-25 association. Structure-function analyses indicate that the C-terminal polybasic region contributes to tomosyn's inhibitory function. These results reveal that tomosyns regulate synaptic transmission by cooperating with synaptobrevin-2/VAMP2 to prevent SNAP-25 binding during SNARE assembly, thereby limiting initial synaptic strength and equalizing it during repetitive stimulation.

Electrophysiological correlates of self-related processing in adults with autism.

The term "self-bias" refers to the human propensity to prioritize self- over other-related stimuli and is believed to influence various stages of the processing stream. By means of event-related potentials (ERPs), it was recently shown that the self-bias in a shape-label matching task modulates early as well as later phases of information processing in neurotypicals. Recent claims suggest autism-related deficits to specifically impact later stages of self-related processing; however, it is unclear whether these claims hold based on current findings. Using the shape-label matching task while recording ERPs in individuals with autism can clarify which stage of self-related processing is specifically affected in this condition. Therefore, this study sought to investigate the temporal course of self-related processing in adults with and without autism. Thirty-two adults with autism and 27 neurotypicals completed a shape-label matching task while ERPs were concomitantly recorded. At the behavioral level, results furnished evidence for a comparable self-bias across groups, with no differences in task performance between adults with and without autism. At the ERP level, the two groups showed a similar self-bias at early stages of self-related information processing (the N1 component). Conversely, the autism group manifested a lessened differentiation between self- and other-related stimuli at later stages (the parietal P3 component). In line with recent claims of later phases of self-related processing being altered in autism, we found an equivalent self-bias between groups at an early, sensory stage of processing, yet a strongly diminished self-bias at a later, cognitive stage in adults with autism.

The relative effectiveness of conditioning one or two attributes to a brand.

Attribute conditioning refers to the phenomenon that target stimuli acquire specific attributes by pairing them with stimuli possessing these attributes. We apply attribute conditioning to a marketing context where brands are often displayed with stimuli possessing semantic attributes to establish brand-attribute associations. In particular, we examine whether it is more effective from a brand image perspective to associate a brand with only one attribute, two related attributes, or two unrelated attributes. Across four experimental studies, we find that pairing a single attribute (e.g., athletic) with a brand is most effective for building brand-attribute associations and that pairing multiple, related attributes (athletic and healthy) is more effective than pairing multiple, unrelated attributes (athletic and smart). Supplementing this finding, an analysis of observational data from real brands suggests that attributing two orthogonal attributes to a brand is associated with negative effects on marketing-relevant outcomes. Our findings extend previous research on multiattribute conditioning and highlight the importance of the number and relationship between attributes for building effective brand associations. (PsycInfo Database Record (c) 2024 APA, all rights reserved).