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Single-cell genomics is a powerful tool for studying heterogeneous tissues such as the brain. Yet little is understood about how genetic variants influence cell-level gene expression. Addressing this, we uniformly processed single-nuclei, multiomics datasets into a resource comprising >2.8 million nuclei from the prefrontal cortex across 388 individuals. For 28 cell types, we assessed population-level variation in expression and chromatin across gene families and drug targets. We identified >550,000 cell type-specific regulatory elements and >1.4 million single-cell expression quantitative trait loci, which we used to build cell-type regulatory and cell-to-cell communication networks. These networks manifest cellular changes in aging and neuropsychiatric disorders. We further constructed an integrative model accurately imputing single-cell expression and simulating perturbations; the model prioritized ~250 disease-risk genes and drug targets with associated cell types.

Few neuropsychiatric disorders have replicable biomarkers, prompting high-resolution and large-scale molecular studies. However, we still lack consensus on a more foundational question: whether quantitative shifts in cell types-the functional unit of life-contribute to neuropsychiatric disorders. Leveraging advances in human brain single-cell methylomics, we deconvolve seven major cell types using bulk DNA methylation profiling across 1270 postmortem brains, including from individuals diagnosed with Alzheimers disease, schizophrenia, and autism. We observe and replicate cell-type compositional shifts for Alzheimers disease (endothelial cell loss), autism (increased microglia), and schizophrenia (decreased oligodendrocytes), and find age- and sex-related changes. Multiple layers of evidence indicate that endothelial cell loss contributes to Alzheimers disease, with comparable effect size to APOE genotype among older people. Genome-wide association identified five genetic loci related to cell-type composition, involving plausible genes for the neurovascular unit (P2RX5 and TRPV3) and excitatory neurons (DPY30 and MEMO1). These results implicate specific cell-type shifts in the pathophysiology of neuropsychiatric disorders.

Abstract Introduction Patient oriented research (POR) invites patients to partner with researchers, clinicians, and other stakeholders, incorporating diverse perspectives to generate scientific evidence meaningful to all parties involved. We adopted a POR approach for this study evaluating the feasibility of conducting a randomized control trial of a novel tri-compartment offloader brace for knee osteoarthritis. We involved patients as partners to enhance study design, implementation and interpretation of key outcomes. Approach Patient involvement consisted of two patient leaders and five patient advisors. Patients participated in 2 virtual focus groups to discuss study outcomes, protocol, results and knowledge translation. Patients were involved in all aspects of the research cycle. Outcomes Patient feedback resulted in changes to study design, documentation, participant recruitment, data collection, results interpretation and knowledge dissemination, improving the participant experience and aligning study outcomes with patient priorities. Study participants showed a high level of protocol adherence and follow-up rates were excellent. We experienced several unexpected benefits including genuine friendships, a deeper understanding of the patient experience, a more pragmatic approach to clinical research, and leadership opportunities for patients. Recommendations We agreed on POR “non-negotiables” to ensure a positive experience for everyone, including creating a safe and comfortable environment, being genuinely receptive to patient feedback, and providing appropriate supports for patients. We strongly recommend that researchers (1) involve patients as early as possible, (2) provide ample and equal opportunities for all patients to be involved, and (3) address system hierarchy by involving patients as equals and fully considering all patient ideas from the beginning of the project. Conclusions While POR is a learning process that is often more challenging than the traditional clinical research approach, the benefits are well worth the additional time and effort required to do it well. Over time, our team experienced a cultural shift and evolution from a top-down research approach to a more inclusive approach considering patient voices as equal to those of researchers. Patient involvement in all aspects of the research process, from question development to results interpretation and dissemination is integral to clinical research advancing equitably.

SUMMARY: There is a demand for novel pathogen mitigation strategies in antibiotic-free poultry production, which has triggered the development of various probiotic application methods. One application method is the in ovo (in egg) administration of probiotics on incubation day (d) 18, where the unhatched chick consumes a probiotic before hatch and prior to being exposed to environmental pathogens. Therefore, the objective of this study was to evaluate the influence of in ovo L. animalis, B. licheniformis, or L. animalis + B. licheniformis (106 cfu/50 μL each) administration on hatch performance (N = 21 egg flats/treatment, 30 eggs/flat), live performance (N = 12 pens/treatment), and d of hatch immuno-physiological parameters (N = 12 birds/treatment) among Ross 708 broiler chicks. Data were analyzed by 1-way ANOVA with significance established at P ≤ 0.05. All in ovo probiotic treatments had greater % hatch than the control treatment, and eggs in ovo-administered with B. licheniformis had lower % late dead and % culled-pipped eggs. However, all in ovo probiotic treatments had lower average chick weights compared to those of the HVT vaccine control group. At hatch, chicks administered with B. licheniformis or L. animalis + B. licheniformis had greater relative ceca weight and lower peripheral blood leukocyte levels compared to those belonging to the control group. There were no effects on live performance during the first 2 wk post-hatch. These results show that B. licheniformis and L. animalis are effective candidates for in ovo probiotic administration, but further investigations regarding immuno-physiological responses to in ovo B. licheniformis administration are necessary.

Squamous cell carcinomas (SCCs), including lung, head & neck, bladder, and skin SCCs often display constitutive activation of the KEAP1-NRF2 pathway. Constitutive activation is achieved through multiple mechanisms, including activating mutations in NFE2L2 (NRF2). To determine the functional consequences of Nrf2 activation on skin SCC development, we assessed the effects of mutant Nrf2E79Q expression, one of the most common activating mutations in human SCCs, on tumor promotion and progression in the mouse skin multistage carcinogenesis model using a DMBA-initiation/TPA-promotion protocol where the Hras A->T mutation (Q61L) is the canonical driver mutation. Nrf2E79Q expression was temporally and conditionally activated in the epidermis at two stages of tumor development: 1) after DMBA initiation in the epidermis but before cutaneous tumor development and 2) in pre-existing DMBA-initiated/TPA-promoted squamous papillomas. Expression of Nrf2E79Q in the epidermis after DMBA initiation but before tumor occurrence inhibited the development/promotion of 70% of squamous papillomas. However, the remaining papillomas often displayed non-canonical Hras and Kras mutations and enhanced progression to SCCs compared to control mice expressing wildtype Nrf2. Nrf2E79Q expression in pre-existing tumors caused rapid regression of 60% of papillomas. The remaining papillomas displayed the expected canonical Hras A->T mutation (Q61L) and enhanced progression to SCCs. These results demonstrate that mutant Nrf2E79Q enhances the promotion and progression of a subset of skin tumors and alters the frequency and diversity of oncogenic Ras mutations when expressed early after initiation.

SUMMARY: This study evaluated the effects of reducing total amino acid (AA) & crude protein (CP) level and supplementation of protease on egg production, egg quality, AA digestibility and economics of egg production in W-36 laying hens from 50-70 wk of age. Five hundred seventy-six Hy-Line W-36 hens were housed in 192 cages consisting of 12 reps of 2 side-by side cages containing 3 hens. Each replicate was fed one of 8 diets in a 4 × 2 factorial arrangement of AA&CP levels (85, 90, 95, and 100% of the breeder recommendations) and protease (exclusion or inclusion). Protease was incorporated into the feed at the dosage recommended by the manufacturer, which is 60 grams per metric ton. Corn and soybean meal-based diets were formulated to meet 100% of the breeder's recommendations for the standardized ileal digestibility of Lys, Met, Thr, Trp, TSAA, Ile, and Val. Our study explored the effects of varying levels of AA&CP set at 85, 90, 95, and 100% of the recommendations for the ileal digestibility of Lys, Thr, Trp, TSAA, Ile, and Val, in diets with and without protease supplementation. All diets were supplemented with phytase at 500 phytase units/kg. Data were analyzed using PROC GLM of SAS 9.4 (2019) and the means were separated using Tukey's multiple comparison tests (P < 0.05). Overall, an interaction was observed between AA&CP level and protease supplementation on egg weight (EW; P = 0.003). The main effect of AA&CP level was observed on haugh unit (HU; P < 0.001) and feed cost (P < 0.001) where higher values were observed in 100 and 95% CP level diets as compared to 90 and 85% CP level diets. Similarly, a decrease in the AA&CP level increased the apparent ileal digestibility (AID) of CP (P < 0.001), Lys (P < 0.001), Met (P < 0.001), Thr (P < 0.001), Trp (P < 0.001), Ile (P < 0.001), and Val (P < 0.001). Protease supplementation increased the AID of CP (P = 0.003), Lys (P = 0.001), Thr (P = 0.009), Ile (P < 0.001), and Val (P < 0.001). In conclusion, lower levels of AA&CP supplemented with protease did not negatively affect the egg production, albumen quality and the cost of production in low AA&CP corn and soybean meal-based diets in the Hy-Line W-36 laying hen.

BackgroundAdherence to clinical practice guidelines for alcohol and drug screening, brief intervention, and referral to treatment (SBIRT) is often inadequate. Mobile apps developed as clinical translation tools could improve the delivery of high fidelity SBIRT.Methods: This study tested the effectiveness of an SBIRT mobile app conceptually aligned with the Theory of Planned Behavior (TPB) to support SBIRT delivery by health care trainees (nursing, social work, internal medicine, psychiatry, and psychology) working in clinical settings (N = 101). Bivariate analyses examined the rate of SBIRT delivery between trainees assigned to the experimental (app) and control (no app) study conditions; as well as the relationship between TPB-based constructs, intention to deliver SBIRT, and screening rates.Results: No significant differences were identified between the study conditions in SBIRT delivery. Significant correlations were found between intent to screen and TPB variables including attitudes/behavioral beliefs concerning substance use treatment (r = .49, p = .01); confidence in clinical skills (r = .36, p = .01); subjective norms (r = .54, p = .01) and perceived behavioral control over appointment time constraints (r = .42, p = .01). Also significant were correlations between percent of patients screened and confidence (r = .24, p = .05); subjective norms (r = .22, p = .05) and perceived behavioral control (r = .28, p = .01).Conclusions: The negative results of the study condition comparisons indicate the need for further investigation of strategies to optimize mobile app utilization, engagement, and effectiveness as a clinical translation tool. Findings of significant correlations between substance use screening rates and both norms and confidence support the potential value of the TPB model in explaining behavior of health care learners in SBIRT delivery.

Neuropsychiatric disorders classically lack defining brain pathologies, but recent work has demonstrated dysregulation at the molecular level, characterized by transcriptomic and epigenetic alterations1-3. In autism spectrum disorder (ASD), this molecular pathology involves the upregulation of microglial, astrocyte and neural-immune genes, the downregulation of synaptic genes, and attenuation of gene-expression gradients in cortex1,2,4-6. However, whether these changes are limited to cortical association regions or are more widespread remains unknown. To address this issue, we performed RNA-sequencing analysis of 725 brain samples spanning 11 cortical areas from 112 post-mortem samples from individuals with ASD and neurotypical controls. We find widespread transcriptomic changes across the cortex in ASD, exhibiting an anterior-to-posterior gradient, with the greatest differences in primary visual cortex, coincident with an attenuation of the typical transcriptomic differences between cortical regions. Single-nucleus RNA-sequencing and methylation profiling demonstrate that this robust molecular signature reflects changes in cell-type-specific gene expression, particularly affecting excitatory neurons and glia. Both rare and common ASD-associated genetic variation converge within a downregulated co-expression module involving synaptic signalling, and common variation alone is enriched within a module of upregulated protein chaperone genes. These results highlight widespread molecular changes across the cerebral cortex in ASD, extending beyond association cortex to broadly involve primary sensory regions.

Some individuals with autism spectrum disorder (ASD) carry functional mutations rarely observed in the general population. We explored the genes disrupted by these variants from joint analysis of protein-truncating variants (PTVs), missense variants and copy number variants (CNVs) in a cohort of 63,237 individuals. We discovered 72 genes associated with ASD at false discovery rate (FDR) ≤ 0.001 (185 at FDR ≤ 0.05). De novo PTVs, damaging missense variants and CNVs represented 57.5%, 21.1% and 8.44% of association evidence, while CNVs conferred greatest relative risk. Meta-analysis with cohorts ascertained for developmental delay (DD) (n = 91,605) yielded 373 genes associated with ASD/DD at FDR ≤ 0.001 (664 at FDR ≤ 0.05), some of which differed in relative frequency of mutation between ASD and DD cohorts. The DD-associated genes were enriched in transcriptomes of progenitor and immature neuronal cells, whereas genes showing stronger evidence in ASD were more enriched in maturing neurons and overlapped with schizophrenia-associated genes, emphasizing that these neuropsychiatric disorders may share common pathways to risk.

SUMMARY: As hens age, egg production and quality decline. Producers are interested in extending the production of their hens to help with production costs and demand. The objective of this study was to investigate the role of the ratio of fine (F) and coarse (C) limestone and exogenous phytase at standard and superdosing levels on performance and egg quality of post-peak hens from 40 to 60 wk of age. A total of 560 Hy-Line W-36 hens (20 replicates of 4 hens per treatment) were randomly assigned into 7 experimental diets in a 2 × 3 + 1 factorial arrangement. The positive control (PC) was formulated to an industry standard that consisted of a 40F:60C limestone ratio without phytase and contained 4.4% calcium (Ca) and 0.44% available phosphorus (avP) for both layer 2 and layer 3 diets. A common negative control (NC) diet was created by reducing Ca and avP by 0.19% and 0.17% respectively compared with the PC. The factorial treatments consisted of 2 limestone ratios (40F:60C and 15F:85C) and 3 Escherichia coli-derived phytase levels (0, 400, and 1,500 FTU/kg) formulated from the NC. Results indicated that 40F:60C at 0 and 400 FTU/kg increased hen-day egg production (HDEP). Limestone ratios of 15F:85C decreased feed intake (FI), increased both eggshell thickness (ST) and weight (SW), but also increased unsaleable eggs (UE) compared to 40F:60C. Additionally, 0 and 400 FTU/kg increased egg quality parameters such as Haugh unit (HU) and albumen height (AH) but 1,500 FTU/kg increased inositol phosphate breakdown as well as bone quality.

INTRODUCTION: Faculty development focused on interprofessional education (IPE) is essential to any IPE initiative aiming to produce a collaborative practice-ready workforce. Many faculty have not received IPE in their own training and struggle with interprofessional teaching. METHODS: To train faculty to conduct a peer-teaching observation and provide feedback focused on interprofessional teaching, we created a 3-hour didactic and skills practice workshop. The didactic portion considered ways interprofessional teaching differed from uniprofessional teaching, discussed elements of effective feedback, and reviewed the critical steps of a peer-teaching observation. In the skills practice portion, participants watched videos of different teaching scenarios and role-played as a peer observer providing feedback to the instructor in the videos. Participants completed a pre/post self-assessment and workshop evaluation form. RESULTS: Eighteen faculty from four professions (dentistry, medicine, nursing, and pharmacy) participated in the workshop from 2020 to 2021. On a 5-point scale (1 = poor, 5 = excellent), participants rated the overall workshop quality 4.9 and the likelihood of making a change in their teaching/professional practice 4.8. Workshop participants self-reported ability to provide feedback to a peer on their interprofessional teaching improved after workshop participation (preworkshop M = 2.9, postworkshop M = 3.8, p < .01). DISCUSSION: This IPE-focused faculty development workshop allows participants to practice skills and share their own interprofessional teaching insights and challenges. The workshop is adaptable for different professions and settings and for in-person or online implementation. It also can be integrated into an existing program or utilized as a stand-alone workshop.

ABSTRACT: Previous work has shown that dietary treatments affect woody breast (WB) incidence differently, which indicates that gut conditions such as gut barrier function, inflammation, and oxidative stress are likely related to WB. In this study, dietary supplementation with antibiotics (bacitracin) or probiotics (Bacillus subtilis) was investigated for their effects on the expression of transcripts related to gut barrier function, inflammation, and oxidative stress in the mucus lining of the jejunum from broilers with or without WB. A split-plot experimental design was used in this study. The dietary treatments served as the main plot factor and the breast muscle condition was the subplot factor. On d 41, jejunum mucus was collected from 1 bird from each of 3 replicate pens in each 3 dietary treatment groups that exhibited WB and an additional bird that contained a normal breast (3 biological replicates/treatment/phenotype; 3 × 3 × 2, total N = 18). Total RNA was extracted using a commercial RNA extraction kit. The expression levels of CLDN1, MUC6, TLR2A, TLR2B, TLR4, IFN-γ, IL-1β, IL-8L1, IL-10, NOS2, and SOD were determined using 2-step RT-qPCR analysis. The gene expression difference in ΔCt values was determined after normalizing with the chicken 18S rRNA gene. When the significant differences occurred between treatments, the relative fold change was calculated using the ΔΔCt method and the significance level was calculated. The PROC GLM procedure of SAS 9.4 was used, and the level of significance was set at P ≤ 0.05. There were no significant interactive effects between diet and the breast muscle condition on the expression of any of the genes tested. However, birds with WB exhibited higher MUC6 (P < 0.0001) gene expression levels than birds with normal breast muscles. In addition, the expression of SOD decreased in birds that were fed the antibiotic diet when compared to birds that were fed the probiotic diet (P = 0.014). In conclusion, WB identified in broilers tested in the current study is attributed to increased expression of mucin, indicating a correlation between WB incidence and gel-forming mucin secretion and pathogen signaling.

Neurodegenerative and neuropsychiatric disorders are pervasive and debilitating conditions characterized by diverse clinical syndromes and comorbidities, whose origins are as complex and heterogeneous as their associated phenotypes. Risk for these disorders involves substantial genetic liability, which has fueled large-scale genetic studies that have led to a flood of discoveries. In turn, these discoveries have exposed substantial gaps in our knowledge with regards to the complicated genetic architecture of each disorder and the substantial amount of genetic overlap among disorders, which implies some degree of shared pathophysiology underlying these clinically distinct, multifactorial disorders. Understanding the role of specific genetic variants will involve resolving the connections between molecular pathways, heterogeneous cell types, specific circuits and disease pathogenesis at the tissue and patient level. We consider the current known genetic basis of these disorders and highlight the utility of molecular systems approaches that establish the function of genetic variation in the context of specific neurobiological networks, cell-types, and life stages. Beyond expanding our knowledge of disease mechanisms, understanding these relationships provides promise for early detection and potential therapeutic interventions.

SUMMARY: To reduce pathogenic bacteria such as Salmonella in feed and ensure a high-quality pellet, diets for broilers, turkeys, and breeders may be pelleted using high conditioning temperatures and extended times. However, increased conditioning temperature and time may alter amino acid conformation regarding secondary and tertiary structure, decrease digestibility and subsequent bird performance, and necessitate increased amino acid formulation density. The objective of this study was to evaluate increased conditioning temperature and exposure time within the pelleting process using a hygienizer (hygienic pelleting) in diets that vary in digestible lysine and corresponding amino acid ratio on Hubbard × Ross 708 starter broiler performance, apparent ileal amino acid digestibility (AIAAD), and requirement. A 5 × 2 factorial arrangement of treatments with 5 levels of digestible lysine (−20, −10%, 2019 Ross Broiler Starter Nutrition recommendation, +10, and +20%) and 2 techniques of pelleting (standard and hygienic) was utilized in a randomized complete block design. Standard pelleting utilized 77°C conditioning for 30 s and hygienic pelleting utilized 88°C conditioning for 60 s and 6-min retention time in a hygienizer. Each treatment was fed to 12 replicate pens of 10 male broiler Hubbard × Ross 708 chicks for a 21-day period. Standard pelleting and hygienic pelleting demonstrated a 1.19 and 1.37% digestible Lys requirement, respectively, using the 2 straight line, 1-breakpoint broken-line regression model. Performance data, AIAAD, and the broken-line regression model suggest an approximate 10% increase in digestible Lys and corresponding amino acid ratio relative to 2019 Ross Broiler Starter Nutrition Specifications for Hygienic Pelleting. Nutritionists should be cognizant that nutritional detriment may accompany procedures used to create hygienic pelleted feed.

Sleep following learning facilitates the consolidation of memories. This effect has often been attributed to sleep-specific factors, such as the presence of sleep spindles or slow waves in the electroencephalogram (EEG). However, recent studies suggest that simply resting quietly while awake could confer a similar memory benefit. In the current study, we examined the effects of sleep, quiet rest, and active wakefulness on the consolidation of declarative and procedural memory. We hypothesized that sleep and eyes-closed quiet rest would both benefit memory compared with a period of active wakefulness. After completing a declarative and a procedural memory task, participants began a 30-min retention period with PSG (polysomnographic) monitoring, in which they either slept (n = 24), quietly rested with their eyes closed (n = 22), or completed a distractor task (n = 29). Following the retention period, participants were again tested on their memory for the two learning tasks. As hypothesized, sleep and quiet rest both led to better performance on the declarative and procedural memory tasks than did the distractor task. Moreover, the performance advantages conferred by rest were indistinguishable from those of sleep. These data suggest that neurobiology specific to sleep might not be necessary to induce the consolidation of memory, at least across very short retention intervals. Instead, offline memory consolidation may function opportunistically, occurring during either sleep or stimulus-free rest, provided a favorable neurobiological milieu and sufficient reduction of new encoding.

Somatostatin interneurons are the earliest born population of cortical inhibitory cells. They are crucial to support normal brain development and function; however, the mechanisms underlying their integration into nascent cortical circuitry are not well understood. In this study, we begin by demonstrating that the maturation of somatostatin interneurons in mouse somatosensory cortex is activity dependent. We then investigated the relationship between activity, alternative splicing, and synapse formation within this population. Specifically, we discovered that the Nova family of RNA-binding proteins are activity-dependent and are essential for the maturation of somatostatin interneurons, as well as their afferent and efferent connectivity. Within this population, Nova2 preferentially mediates the alternative splicing of genes required for axonal formation and synaptic function independently from its effect on gene expression. Hence, our work demonstrates that the Nova family of proteins through alternative splicing are centrally involved in coupling developmental neuronal activity to cortical circuit formation.

SUMMARY: The objective of this study was to evaluate the effect of feeding varying amino acid densities (AAD) and AME levels during the finisher phase (d 28–41) on male Cobb MV × Cobb 500 broiler performance, processing, and economics. A 3 AAD (Low-AAD, Medium (Med)-AAD, and High-AAD) × 4 AME (Low-AME, Med-AME, High-AME, and Very High-AME) factorial arrangement was utilized. Feeding Med-AAD or High-AAD maximized d 28 to 40 BW gain (BWG) regardless of AME level fed. As AAD increased, a stepwise reduction in FCR and ADFI was observed from d 28 to 40. For the main effect of AME, broilers fed Very High-AME improved FCR from d 28 to 40, with those fed Low-AME having a higher FCR and FCR of birds fed Med-AME and High-AME being intermediate. Birds fed Low-AME increased ADFI throughout but were similar to Med-AME from d 28 to 40. An interaction was detected for carcass yield wherein broilers fed Med-AAD + Low-AME maximized carcass yield. Feeding High-AAD maximized breast and tender yield and reduced fat pad yield. For the main effect of AME, fat pad yield was greatest in birds fed High-AME and Very High-AME. Overall, increasing d 28 to 41 AAD improved performance and processing variables; Very High-AME level improved FCR and ADFI. These data demonstrate the importance of evaluating the response of a particular broiler strain to varying AAD and AME levels, as these nutrients can impact performance and ultimately producer profitability.

SUMMARY: A previous companion study utilized a 2 GS (Fast Growing [FG] or High Yielding [HY]) × 2 FF (Crumbles [C] or Intact Pellets [IP]) × 3 FQ (Low, Medium, or High) factorial arrangement to determine their impact on broiler performance. The 2 FF, C or P, were fed in the starter diet (d 0–18) as one of 3 FQ: low-2,210 µm (C) or 40% intact pellets (IP), medium-3,010 µm (C) or 60% IP, and high-3388 µm (C) or 80% IP. From 0 to 18 d, feeding increased FQ improved overall BW and BWG. The current study determined the APSC from d 0 to 6 and d 0 to 18 among these GS, FF, and FQ. This study utilized a subsample of pens from the companion study. From d 0 to 6 and 0 to 18, HY GS had a higher APSC. At d 0 to 18, GS × FF interacted, demonstrating that APSC only differed between GS when pellets were fed, with HY consuming a larger size. A FF × FQ interaction on 0–6 and 0–18 d demonstrated that, regardless of FF broilers fed high FQ, had larger APSC. For birds fed low and medium FQ as a C, APSC was larger, but smaller when fed as IP. Correlations were found showing that as APSC increased, d 0 to 6 BW, BW gain, and FCR improved; this also occured for d 0 to 18 FCR. Data suggest that APSC is dependent upon GS and FF presented; further research is needed to verify APS preference when provided ad libitum amounts of each particle size.

Recent studies demonstrate that eyes-closed rest benefits memory consolidation, perhaps due to reduced attention to environmental stimuli. Here, we asked whether focusing attention to "internal" thoughts and feelings after learning similarly blocks memory consolidation. Verbal memory was tested following an eyes-closed consolidation period filled with either focused attention to breath or quiet rest. Although breath-focus did not impair memory relative to quiet rest overall, participants who reported being more successful in maintaining breath-focus during this condition showed increased forgetting. We interpret these findings as incompatible with a simple sensory-interference-based account of rest's effect on memory.