Journal of Neurogenetics

Objectives: To determine whether distinct anxiety phenotypes (rumination, global anxiety and worry; RAW) are associated with plasma brain-derived (BD) phosphorylated-tau (p-tau) biomarkers of Alzheimer's disease (AD) in older adults. Design: Cross-sectional observational study. Methods: Participants were 89 older adults aged 50[≥]years (mean age 64.3{+/-}8.8 years) enrolled in the RAW Brain study (R01 MH108509) in Pittsburgh, Pennsylvania. Anxiety phenotypes were assessed using the Penn State Worry Questionnaire (PWSQ[≥]50), Rumination Subscale from the Response Style Questionnaire (RSQ>50), and Hamilton Anxiety Rating Scale (HARS>17). Participants were classified as High-RAW if they exceeded established severity thresholds on [≥]1 domain; continuous anxiety scores were used for primary analyses. Plasma BD-tau, BD-p-tau181, BD-p-tau217, and BD-p-tau231, and matched non-BD versions were quantified using the NULISAseq CNS Panel. Associations between anxiety phenotypes and tau biomarkers were examined using multivariable linear regression adjusted for age, sex, and APOE4 status, with Benjamini-Hochberg false discovery rate correction. Results: Greater global anxiety was associated with higher plasma BD-p-tau181 ({beta}=0.308, 95% CI [0.003,0.048], p=0.047). This association was also observed categorically (adjusted Cohen's d=0.462, p=0.049). Worry and rumination were not associated with BD-p-tau181, and no anxiety phenotypes were associated with BD-tau, BD-p-tau217, BD-p-tau231, or corresponding non-BD alternatives. Conclusions: Greater late-life global anxiety, but not worry or rumination, was selectively associated with higher plasma BD-p-tau181. These findings extend epidemiologic evidence linking anxiety to AD risk by identifying a biologically specific blood-based correlate linking anxiety with AD-related p-tau biology. Longitudinal studies incorporating amyloid and tau biomarker confirmation are needed to determine whether anxiety precedes, accompanies, or results from early AD pathophysiology.

Transgenic GAL4 lines are widely used in Drosophila to label and manipulate specific cell types, but the resulting reporter expression patterns do not always represent ongoing driver activity. We generated a collection of neuropeptide T2A-GAL4 knock-ins and characterized their expression by native-fluorescence imaging using UAS-CD8::GFP and a Brp-miRFP680 knock-in, eliminating the need for immunostaining. During characterization of these lines, we observed cases in which adult GFP patterns did not match the on-going GAL4 activity, as well as cases in which GAL4 produced adverse cellular effects. Using temperature-sensitive GAL80, we found that adult GFP expression in certain neurons driven by CCHa2-T2A-GAL4 and Dsk-T2A-GAL4 depended on GAL4 activity before eclosion and was not reproduced by adult-only induction. Several GAL4 lines also showed dominant developmental phenotypes. Heterozygous Burs-T2A-GAL4 animals, for example, displayed wing-expansion defects that were fully suppressed by GAL80. Severe wing defects were not accompanied by a reduction in the number of reporter-labeled neurons before eclosion, but the morphology of these neurons were affected. These observations illustrate how developmental driver history and GAL4-associated cellular effects can influence reporter patterns in the adult nervous system.

The sexual identity of specific tissues can be altered through cell-autonomous manipulation of the sex determination pathway in the nematode Caenorhabditis elegans. Previously described sexually mosaic strains carried multicopy transgenic arrays that are potentially subject to transgene silencing or misexpression artefacts. We used here the CRISPR/Cas9 system to engineer strains in which coding sequences for either a masculinizing agent (FEM-3) or a feminizing agent (intracellular domain of TRA-2) are inserted with a trans-splicing system at the 3 end of endogenous genetic loci that are known to be expressed in either a neuron-specific (rab-3) or gut-specific (elt-2) manner. We validate the tissue-specific sex-transformations induced by these genomic manipulations through expression analysis of known sex-specific genes and through assaying behavioral readouts. These strains may serve the worm community as standard reference tools to study the consequences of tissue-specific sex transformations.

Objectives: The rising prevalence of financial exploitation among older adults has prompted research on the cognitive mechanisms underlying financial decision making in later life. Prior work has demonstrated a link between cognitive impairment and financial vulnerability. The present study examined the unique contributions of composite measures of memory and executive functioning to scam susceptibility in middle-aged to older adults without dementia. Methods: One hundred and fifty-one participants (age= 68.2 {+/-} 8.0 years, 72.2% female, years of education= 16.4 {+/-} 2.2) completed standard neuropsychological measures from the Uniform Data Set (UDS), Version 3, and the California Verbal Learning Test, Second Edition, and a self-report scam susceptibility measure. Psychometrically robust composite scores, the UDS-M+ and UDS3-EF, were computed to represent memory and executive functioning domains. Linear regression models examined independent associations between UDS-M+ and UDS3-EF performance and scam susceptibility. Nested model comparisons evaluated unique contributions of the UDS-M+ and UDS3-EF when included in a joint model. Results: Lower UDS-M+ (b= -0.25, 95% CI [-0.45, -0.04], p= .018) and lower UDS3-EF (b= -0.33, 95% CI [-0.54, -0.12], p= .002) scores were independently associated with greater scam susceptibility after adjusting for age, sex, years of education, and income. However, only UDS3-EF remained significant in a joint model. Post hoc analyses revealed that only memory was associated with scam susceptibility in middle-aged adults whereas only executive functioning was significant in older adults. Conclusions: These findings highlight the importance of monitoring memory and executive functioning across middle to older adulthood when considering susceptibility to scams.

OBJECTIVE: Research on neighborhood context and cognitive health often relies on composite indices that collapse neighborhood socioeconomic status (SES) and aspects of the built environment into a single score. This study examined the extent to which neighborhood SES and built environment were associated with late-life cognitive impairment. METHOD: Participants were 281 predominantly Black older adults who completed the Self-Administered Gerocognitive Exam (SAGE), a brief cognitive screening measure. Participants also provided their home address to obtain four neighborhood metrics: Two composite measures of neighborhood disadvantage, the Environmental Justice Index (EJI) and the EJI Social-Environmental Rank, as well as the EJI Social Vulnerability Rank as a measure of neighborhood SES, and the EJI Environmental Burden Rank to measure exposure to hazardous aspects of the built environment. RESULTS: Lower neighborhood SES was associated with higher odds of cognitive impairment. In contrast, greater environmental burden was associated with lower odds of impairment, with years of education as a moderator; the association was strongest among individuals with high levels of education. The two composite scores were not associated with cognitive impairment. CONCLUSION: Composite measures of neighborhood features may obscure distinct pathways of their effects on cognition.

Site-specific recombinases, such as Cre recombinase (Cre) and FLP recombinase (FLPase), are popular tools that enable precise spatiotemporal control over fluorescent reporter genes. However, studies show that increased Cre expression can lead to cytotoxic phenotypes including DNA damage, cell cycle arrest, cell death, and aneuploidy. In contrast, there is little-to-no known cytotoxicity associated with FLPase. FLPase is considered to be the safer, if less efficient, recombinase. To compensate for decreased efficiency, researchers will often increase expression of the FLPase to ensure a complete gene edit. Despite previous evidence that FLPase is nontoxic at high doses, we report that high expression of FLPase from multi-copy extrachromosomal arrays can disrupt synaptic protein trafficking in C. elegans. We discovered several off-target effects of high FLPase expression including mislocalization of the pre-synaptic marker FLPon::mScarlet::SYD-2 and decreased synaptic abundance, transport, and expression of the ionotropic glutamate receptor subunit GLR-1. Neither overexpression of an inactive FLPase, low activity FLPase, nor mCherry under the same promoter elicited these effects. However, single-copy expression of the FLPase construct reliably tags FLPon::mScarlet::SYD-2 and does not lead to these off-target effects. This work supports growing awareness that tyrosine recombinases can become cytotoxic at high doses.

Chemotaxis assays are widely used to quantify sensory-driven behavioral responses in Caenorhabditis elegans. Sodium azide is commonly used to immobilize animals at assay endpoints, although concerns have been raised that diffusion beyond the endpoint region could affect behavior before a choice is completed. We compared sodium azide-mediated immobilization with post-assay cooling using the repellent 1-octanol and attractant diacetyl. Wild-type animals showed robust avoidance of 30% 1-octanol and attraction to 0.5% diacetyl with either method, across multiple assay durations, and with or without worm food Escherichia coli OP50. These results support post-assay cooling as an alternative endpoint-scoring method.

Objectives Alzheimer's disease (AD) progresses from a cognitively unimpaired stage through mild cognitive impairment (MCI) to dementia. Because language impairment often emerges before dementia, it is a candidate marker of disease stage. Yet the standard batteries of large AD research cohorts rely on a narrow set of tests that mainly evaluate the lexical-semantic domain (i.e., picture naming and verbal fluency), leaving other language domains, such as semantic memory, phonology, morphosyntax, and single-word reading, rarely assessed. We evaluated the diagnostic utility of standard and specialized language tests for distinguishing amyloid-positive MCI and dementia of the Alzheimer's type (DAT) from healthy controls (HC) and from each other. Methods We included 286 participants from the National Alzheimer's Coordinating Center (125 HC, 53 MCI, 108 DAT) assessed with three standard tests from UDSv3 (semantic fluency, letter fluency, and Multilingual Naming Test [MINT]) and nine specialized tests from the FTLD Module (regular and irregular word reading, semantic word-picture matching, semantic associates, Northwestern Anagram Test - Short Form [NAT-S], Northwestern Naming Battery noun and verb naming, sentence repetition, and sentence reading). For each test, the area under the receiver operating characteristic curve (AUC) was computed for DAT vs. HC, MCI vs. HC, and DAT vs. MCI, and AUCs were compared between tests with DeLong's test. Results Eight of 12 measures reached acceptable discrimination (AUC [≥] .700) for DAT vs. HC, five for MCI vs. HC, but only semantic fluency and NAT-S for DAT vs. MCI. Semantic fluency showed the highest discrimination in each contrast, though it did not significantly outperform the NAT-S for MCI vs. HC or DAT vs. MCI. MINT, letter fluency, and irregular word reading reached clinically meaningful accuracy in distinguishing MCI and DAT from HC, but not DAT from MCI. The remaining specialized tests reached it only for DAT vs. HC, or not at all. In a post hoc regression, adjusting for cognitive flexibility and working memory attenuated group differences; the DAT difference was no longer significant, whereas the MCI difference remained significant. Discussion Semantic fluency was the strongest single language marker across the AD continuum. Among specialized tests, NAT-S was the most informative, challenging the view that syntax is spared in early AD. Its performance was partly explained by executive function, mainly in DAT, whereas a residual morphosyntactic component remained in MCI, and several specialized tests were limited by ceiling effects. Clinically, the NAT-S may complement semantic fluency for staging, and specialized tests may help characterize heterogeneous language profiles in MCI and DAT.

Intracellular neurofibrillary tangles formed from hyperphosphorylated tau and extracellular amyloid plaques containing aggregated A{beta}-peptides, specific cleavage products of the Amyloid Precursor Protein (APP), are the primary histopathological hallmarks of Alzheimers Disease (AD), the leading cause of dementia in humans. However, the initiating steps that lead to these pathologies and early neurodegeneration, and the mechanisms by which tau- and A{beta}-induced effects might be linked remain unclear. Using the prostate-like secondary cell (SC) in Drosophila, we recently showed that A{beta} modulates normal APP- and membrane-associated protein aggregation in the dense-core granule (DCG) compartments of the regulated secretory pathway by interfering with subsequent membrane:DCG dissociation. This disrupts endolysosomal trafficking and propagates the resulting endolysosomal defects to other cells that endocytose the secreted abnormal DCG proteins. Here we show that overexpressing human tau also disrupts DCG aggregation and membrane:DCG dissociation inside SC secretory compartments, leading to increased endolysosomal targeting of these compartments. In a genetic screen, we find that knockdown of cofilin, which encodes an actin-severing protein required for dynamic remodelling of microfilaments, generates a similar phenotype. Consistent with this, overexpression of Cofilin, which is known to suppress tau-induced neurodegeneration in flies, reduces tau-induced DCG defects in SCs. Indeed, we find that Cofilin overexpression also suppresses A{beta}-induced degeneration in the fly eye. We conclude that membrane:DCG aggregate dissociation in DCG compartments is disrupted by both tau- and A{beta}-induced genetic changes that are relevant to AD, and this partially involves inhibition of actin cytoskeleton dynamics. Increasing actin remodelling activity can suppress neurodegeneration induced by both tau and A{beta}, suggesting that this process provides an important functional link between them that might be targeted therapeutically.

Tardigrades, key ecdysozoan organisms, have poorly characterized daily behavioral rhythms linked to their molting (ecdysial) cycle. We continuously tracked the locomotor activity of the model tardigrade Hypsibius exemplaris using infrared videography and deep learning-based automated tracking under three photic regimes: a light/dark cycle, constant darkness following entrainment, and constant darkness without prior entrainment. We identified a robust endogenous circadian-like rhythm in locomotor activity that persisted in constant darkness, independent of photic input. Although the light/dark cycle enhanced the rhythm amplitude and the overall animal activity, it was not required for rhythmicity. Individual phases varied widely, indicating a lack of population-level synchronization. Furthermore, we developed a behavioral classification system based on activity level, which enables categorizing distinct behaviors. Behavioral patterns were modulated by distinct ecdysial stages, with quiescence gating premolt and time-of-day dependent patterns in feeding and wandering. These findings provide evidence consistent with an endogenous, circadian-like timing system in tardigrades that integrates daily activity with molting cycles and establish H. exemplaris as a promising model for studying ecdysozoan chronobiology.

Attention deficit/hyperactivity disorder (ADHD) is a common neurodevelopmental condition characterized by inattention, impulsivity, and hyperactivity, and is believed to involve altered dopamine signaling. Growing evidence suggests that the gut microbiome may contribute to neurobehavioral conditions, including ADHD. In this study, we examined differences in gut microbiome composition, measured using metagenomics, between healthy and ADHD-affected individuals, and evaluated age- and gender-related variation within the ADHD population. We analyzed publicly available gut microbiome datasets from the American Gut Project, NCBI, and MGnify, and retrieved data from 496 ADHD and 419 healthy individuals. Further, metadata and taxonomic abundance were processed using RStudio, normalized to sequencing depth, and analyzed at the genus level. Both groups shared several gut bacteria, including Blautia, Bacteroides, Faecalibacterium, Roseburia, and Alistipes, indicating a broadly similar core microbiome. However, individuals with ADHD showed differences in the stability and persistence of certain core taxa, such as Proteobacteria and the unclassified genus Prevotellaceae, which were uniquely present in the ADHD group. Further, PERMANOVA showed that age and gender explained a small yet significant difference in microbial variation (Rsquare = 0.0437, p = 0.001), with greater variability observed between males and females than across age groups. Dispersion analysis revealed significant microbial variability between males and females (p = 0.001), whereas age groups showed no significant variation (p = 0.792). Overall, these results suggest that differences in the gut microbiome between individuals with ADHD and healthy individuals are subtle. Clinically, this highlights the need to consider broader biological, environmental, and lifestyle influences- such as diet, medication use, and comorbidities- when evaluating the potential role of the gut microbiomes in ADHD.

A key pathophysiological hallmark of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) is loss of nuclear localization and abnormal cytoplasmic aggregation of TAR DNA-binding protein 43 (TDP-43). Upstream factors that trigger the onset and progression of such neurodegenerative diseases are largely unknown. Aging and environmental factors contribute as potential risk factors. But epidemiological evidence suggests psychological stressors such as social isolation and loneliness are also associated with increased risks of neurodegenerative disease [1-8]. We examined the impacts of social isolation on TDP-43 related neurodegeneration. To investigate the potential for causative impact on neurodegeneration, we compared the effects of social isolation versus social enrichment in Drosophila. We utilized an established social isolation paradigm in which flies were either housed alone or in groups as young adults. We found that early-life social isolation acts as a primer to exacerbate the rate of neurodegeneration in response to subsequent induction of pathological levels of TDP-43. Social isolation stress drives more rapid activation of mdg4, an endogenous retrovirus that functionally mediates TDP-43 effects and more aggressive propagation of TDP-43 protein pathology from surface glial cells to nearby neurons. Shortened life span also ensues. We demonstrate that providing isolated flies with visual, olfactory and tactile social cues from conspecifics living behind a divider was insufficient to alleviate these effects of social isolation on neurodegeneration. Our findings have implications for the association between psychological stressors such as loneliness and risk of neurodegenerative diseases in humans and provide a platform to investigate mechanistic underpinnings in animal models.

Abstract The neurotransmitter serotonin (5-hydroxytryptamine; 5-HT) controls behaviours that are essential for the parasitic lifestyle of plant parasitic nematodes (PPNs). Here, we used serotonin immunochemistry and in situ hybridization for the serotonin synthesis genes, Gpa-tph-1 and Gpa-bas-1, to map its signalling pathways in Globodera pallida J2 infective stages. We identified fourteen serotonin-immunoreactive cell bodies around the pharyngeal median bulb and revealed that eleven of them co-express Gpa-tph-1 and Gpa-bas-1, confirming these are serotonin-synthesising neurons. Serotonin immunoreactivity was also localised in structures consistent with labelling of amphids, and in sixteen non-neuronal cells located in the middle and posterior region of the nematode. Depletion of serotonin with reserpine reduced the serotonin immunoreactivity in all fourteen cell bodies confirming their neuronal identity, but not in non-neuronal tissue. We identified additional, but infrequent expression of Gpa-tph-1 in two cells and expression of Gpa-bas-1 in one pair of symmetrically bilateral cells anterior to the median bulb, that lacked serotonin immunoreactivity. The results reveal distinct pools of serotonin distribution in G. pallida and indicate serotonin signalling is more complex than in other parasitic nematodes and C. elegans, highlighting evolutionary divergence in neurotransmitter identity.

Animal pose estimation typically requires manual annotation of body parts to train neural networks for each new recording system or environmental condition. Here we use a general-purpose vision-language model (VLM) to generate annotations in multi-animal recordings of fruit flies (Drosophila melanogaster). Initial annotations follow written anatomical descriptions and are reviewed by an independent VLM session using neighboring video frames. On 120 held-out fly images, AI annotations had a median distance of 2.28 pixels from human labels, with 84.0% agreement within eight pixels, averaged across landmark types. The comparison included 10 landmarks, with head and eyes analyzed separately because of systematic differences in placement. Review improved agreement primarily at the feet, with the largest remaining discrepancies at positions with limited visual evidence. Visual inspection revealed cases in which the AI annotations were more anatomically accurate than those of an experienced human annotator, particularly at the feet. A convolutional network trained using SLEAP on 300 AI-annotated fly images showed similar agreement with human labels and closely reproduced the AI annotations on held-out images, with a median distance of 1.56 pixels. We also applied the procedure to recordings from another laboratory, using new AI annotations to train a separate pose network. Although VLM annotation required minutes per fly, the trained networks enabled pose estimation throughout more than 700,000 frames. These results demonstrate that VLM annotations can support fast animal pose estimation without manually labeled training images.

Background: Emerging evidence suggested the role of sleep and circadian rhythm in the development of mental resilience, but the association of an integrated analysis of objective sleep and circadian measures with multifaceted mental resilience in healthy adults has been underexplored. Objective: This study aims to explore the associations between multifaceted mental resilience and sleep and circadian rhythm in healthy adults. Methods: A total of 156 healthy adults (Age: Mean (SD): 29.54 (5.28) years; 60.9% female) underwent 14-day actigraphy, one night polysomnography (PSG) and dim light melatonin onset (DLMO) assessments. Resilience was operationalized as both a coping capacity and a positive mental health outcome in the context of accumulative adversity (life stressful events and childhood adversities). We first identified the covariance patterns between multiple sleep and circadian health domains (Sleep macro- and microstructures, dim light melatonin onset, rest activity pattern) and multifaceted mental resilience using partial least squares regression (PLSR), and followed by univariate analysis as complementary to illustrate the distribution trends of sleep and circadian rhythm parameters across integrative resilience score derived from principal component analysis (PCA). Findings: The PLSR model identified a modest but significant association of greater mental resilience with a sleep and circadian profile (Bootstrapping r = 0.15, 95%CI: 0.02-0.28), characterized by greater frontal-central slow waves activity in NREM sleep, greater frontal theta activity in REM sleep, lower occipital spectral power across fast and slow frequency bands in NREM and REM sleep, and probably earlier dim light melatonin onset timing. Conclusions: Sleep and circadian metrics, specifically frontal slow waves and REM theta activity, attenuated overall occipital spectral power during sleep, and earlier dim light melatonin onset time, represent quantifiable targets that may serve to improve resilience and stress-related mental health in adults. Longitudinal and interventional studies are needed to establish whether sleep interventions can be endorsed as evidence-based approaches for resilience enhancement. Clinical implications: Targeting sleep and circadian aspects may be an important approach to promote stress-related mental health.

People from UK South Asian communities experience barriers to accessing dementia care. Evidence suggests that evidence-based psychosocial dementia interventions can be successfully adapted for different cultural contexts, but further evidence is needed on how adaptations are developed and refined in practice. We aimed to adapt GRACE-family (Goal diRected, Accessible and Evidence-based Care), an evidence-based, manualised post-diagnostic dementia intervention, comprising 4-8 sessions for family carers and where possible, people with dementia, for Bengali-speaking communities. In a theory-informed process, a co-adaptation group encompassing lived, academic and clinical/professional experience developed consensus regarding which intervention functions were core. They then explored how adaptation might, while retaining these, account for cultural and linguistic contexts, informed by focus groups with memory service and Bengali community organisation staff (n=14). We developed a prototype adaptation, then piloted it with four Bangladeshi families supporting a person with dementia. Post-intervention, participants were asked about goal attainment, intervention acceptability, and qualitatively interviewed. Pilot data and a focus group with Sylheti-speaking family carers informed further refinements (n=6). Across data sources, two themes informed adaptation: (1) stigma, trust and community networks shaped access to dementia support; (2) flexible delivery was needed to respond to diverse language, communication and cultural needs. 3/4 pilot families completed the prototype intervention, which was in English as initial consultation cautioned against developing a written manual in a predominantly spoken language. Participants reported improvement on 7/8 personalised goals set, and all considered the intervention acceptable. Pilot delivery identified challenges using English-language materials during interpreter-mediated sessions. Iterative refinements added standard Bengali materials and spoken Sylheti audiovisual resources to broaden delivery options and facilitator training for culturally responsive and interpreter-mediated delivery. Our co-adaptation approach enabled stakeholders to advise from different, including experiential perspectives. It may inform future approaches to cultural and linguistically adaptations of psychological interventions for minoritised communities.

Background: Insufficient oxygen (hypoxia) or complete lack of oxygen (anoxia) causes cellular dysfunction and death in terrestrial animals. As a result, animals have developed homeostatic strategies, including changes in movement strategies to escape and survive stressful environments. Caenorhabditis elegans respond to changes in oxygen by increasing the frequency of reversals (backward movement) and increasing locomotor speed; these behaviors enable them to escape unfavorable environments and move to more suitable ones. The molecular and neural mechanisms underlying behavioral responses to low-oxygen environments are incomplete. Neuropeptides regulate C. elegans behaviors by modifying neural circuit function in response to hypoxia. Loss of function in C. elegans neuropeptide nlp-40 and its receptor aex-2 increased survival to anoxia [1]. However, the cells in which NLP-40 and AEX-2 function for anoxia, as well as the mechanisms underlying responses to anoxia, remain elusive. We hypothesized that NLP-40 and AEX-2 regulate anoxia by altering escape behaviors. Results: We found that neuropeptide NLP-40 functions in cholinergic neurons and AEX-2 functions in cholinergic or GLR-1 interneurons or motor neurons to regulate susceptibility to anoxia. We found that loss of either nlp-40 or aex-2 decreased locomotor activity and reversal frequency early during 48 hours of anoxia, suggesting that nlp-40 and aex-2 are required for the proper execution of these behaviors. We found that neuronal knockdown of nlp-40 recapitulated the behaviors observed in nlp-40 mutant animals and increased survival; however, restoring aex-2 function in glr-1 interneurons or motor neurons was sufficient to rescue behaviors to wild-type and restore vulnerability to anoxia. Conclusions: Overall, our results suggest that NLP-40 and AEX-2 likely function in cholinergic or glr-1 interneurons or motor neurons to regulate escape behaviors during anoxia. Escape behaviors are part of the homeostatic response and are typically protective in the wild; however, in our paradigm, we show that loss of either nlp-40 or aex-2 reduces escape responses and confers protection against 48 hours of anoxia. Collectively, these results highlight a new role for NLP-40 and AEX-2 in anoxia and shed light on mechanisms underlying the adaptive responses to anoxia.

Autistic individuals with intellectual disability (ASD+ID) and profound autism are rarely studied in neuroscience research due to barriers including communication challenges, sensory sensitivities, anxiety, and elevated behavioral support needs. Consequently, biomarker development in autism fails to include the portion of the autistic population with highest support needs and greatest potential to benefit from treatments derived from imminent advances in personalized medicine. Here we present a multimodal, participant-centered protocol for simultaneous electroencephalography (EEG) and eye-tracking (ET) acquisition to advance clinical trial readiness research in this underserved population. The protocol combines individualized behavioral supports and an adaptive EEG-ET acquisition platform. Pre-visit caregiver interviews informed participant-specific research plans, including customized visual supports, reinforcement strategies, and EEG net desensitization procedures. A gaze-contingent stimulus presentation system used participant-preferred videos to support attention to experimental stimuli and behavior consistent with data acquisition (e.g., refraining from movement) while dynamically adapting stimulus delivery based on visual engagement with the assay and real-time assessment of data quality. By quantifying attended trials in real time and prioritizing unattended stimulus classes, the paradigm maximizes acquisition efficiency to minimize participant burden. This approach was utilized to deploy candidate biomarkers with demonstrated replicability in the Autism Biomarkers Consortium for Clinical Trials (ABC-CT): the N170 event-related potential to faces, resting-state EEG, and visual attention to human faces (measured with ET). Feasibility was evaluated in 76 children with ASD+ID (M age = 8.76 years; IQ = 32.4) and a small comparison sample of 8 children with intellectual disability without autism (M age = 8.91 years; IQ = 45.9). Usable EEG data (>7 minutes) were acquired in 61% (ASD+ID) and 88% (ID) of participants, respectively. Sufficient ET data were obtained from 67% (ASD+ID) and 63% (ID) of participants, respectively. Findings demonstrate the feasibility of inclusive biomarker acquisition and provide a framework for expanding participation of individuals with ASD+ID and profound autism in neuroscience research. Ongoing research seeks to evaluate this approach in larger samples and to demonstrate feasibility in multisite research studies.

Study Objectives: To characterize neural correlates of memory and emotion reactivity in individuals with insomnia disorder (INS) compared to good sleepers (GS) and to investigate associated short- and long-term changes following cognitive-behavioural therapy for insomnia (CBTi). Methods: This randomized controlled trial included 56 INS and 20 GS. At baseline, participants underwent resting-state and task-based functional magnetic resonance imaging, including working memory, emotion reactivity, and face-name associative memory tasks. They filled out sleep diaries and self-report questionnaires about sleep, cognition, and emotions, and underwent in-lab polysomnography. Subsequently, participants with INS were randomized to either the 3-month treatment group (TX) or wait-list control group (WL). Both TX and WL groups repeated baseline assessments at three months post-randomization and completed sleep diaries and questionnaires one year after finishing CBTi. Results: At baseline, the INS group reported higher insomnia severity, greater subjective cognitive impairment, and perceived sleep-related stimuli as more negative than GS. Contrasting with these subjective evaluations, no group differences were found in memory performance, task-based brain activity, or resting-state functional connectivity. At three months post-randomization and one-year follow-up, CBTi significantly improved subjective, but not objective, cognitive functioning and sleep in adults with INS. No significant treatment-related changes were observed in valence ratings of sleep images, task-based brain activity, or resting-state functional connectivity. Conclusions: Beyond its primary role in improving insomnia symptoms, CBTi may have long-term benefits for individuals with INS who experience concerns about their cognitive functioning, even in the absence of objective changes in neural processing of cognitive and emotional tasks.

Over the course of evolution, many species have adapted to the environmental conditions found at higher latitudes. This is likely also true of the circadian system, which regulates daily behavior and helps measure day length to facilitate adaptation to the seasons, though how the circadian clock adapts to latitude remains largely unknown. Here, we investigated daily activity patterns under long photoperiods and in constant darkness, as well as the underlying circadian clock network, in 52 Drosophila species from around the world. We found a strong, significant positive correlation between the clock's plasticity, measured as the phase difference between morning and evening activity, and the average geographical latitude from which the species originate. The mechanisms underpinning this increase in plasticity differed between species: in the subgenus Siphlodora, the clock weakened with increasing latitude, accompanied by a reduction of the neuropeptide Pigment-Dispersing Factor in the small ventrolateral clock neurons (s-LNv), whereas in the subgenus Drosophila the opposite was true. The subgenus Sophophora, by contrast, showed a negative correlation between circadian plasticity and PDF levels in the s-LNv axons. These results suggest that Drosophila species have evolved diverse, independent strategies to adapt their circadian clock to higher latitudes.