{"artifact":{"id":"experiment_proposal-afa252d5-e3ae-4839-9258-316fe0045c76","artifact_type":"experiment_proposal","entity_ids":"[\"h-bdbd2120\"]","title":"Experiment Proposal: h-bdbd2120","quality_score":0.75,"created_by":"forge_experiment_proposal_generator","provenance_chain":"[]","content_hash":"2f2ed0349a6d725e9cef931479fdec85e33ce9b766b46b200f71d406985d5a65","metadata":{"citations":["31076275","35151204","36450248","27929004","31578527","36879142","37384704","38642614","39964974","33127896"],"generated_at":"2026-04-28T20:00:29.804839+00:00","intervention":"40 Hz audiovisual sensory entrainment delivered via custom Arduino-controlled device: 40 Hz square-wave LED array (luminance 250 cd/m², 50% duty cycle) + 40 Hz click train (2 ms duration, 70 dB SPL) co-administered for 1 hour/day, 7 days/week for 6 weeks; sham group receives identical apparatus with LEDs at 0 Hz and clicks at 0.5 Hz (below gamma range)","key_controls":["APP/PS1ΔE9 sham-stimulated mice (0.5 Hz stimulation, no entrainment) — disease model negative control","C57BL/6J wild-type mice with 40 Hz stimulation — positive control for entrainment efficacy","C57BL/6J wild-type mice with sham stimulation — baseline reference for all readouts","20 Hz entrainment APP/PS1 group (frequency specificity control, per Soula et al. 2023 PMID:36879142)","Entrained tissue collection at 1 hour post-final session (acute molecular readouts) vs 24-hour washout (sustained effect assessment)","Age-matched untreated APP/PS1 mice for longitudinal trajectory comparison"],"model_system":"APP/PS1ΔE9 mice on C57BL/6J background, both sexes, 6 months old at intervention start (moderate Aβ pathology burden), n=30 per group (power 0.80, α=0.05, two-tailed, expected d=0.85 for primary outcome), with 15% attrition buffer resulting in n=35/group; age/sex-matched C57BL/6J wild-type mice (n=20) as positive controls for gamma entrainment response","hypothesis_id":"h-bdbd2120","timeline_weeks":18,"_schema_version":1,"null_hypothesis":"40 Hz audiovisual gamma entrainment (1 hour/day for 6 weeks) does not restore hippocampal-prefrontal cortical synchrony, does not reduce Aβ or tau pathology, does not activate microglial phagocytosis, and does not improve cognitive performance in 6-month-old APP/PS1ΔE9 mice compared to sham-stimulated age-matched controls","primary_readout":"Hippocampal-prefrontal cortical gamma coherence measured via bilateral silicon probe LFP recordings (32 channels) during active stimulation and 30-min post-stimulation; primary endpoint is phase-locking value (PLV) at 35-45 Hz band between CA1 LFP and prelimbic PFC LFP, analyzed with Chronux toolbox","estimated_cost_usd":287500.0,"secondary_readouts":["Aβ40/Aβ42 levels in hippocampus and prefrontal cortex by ELISA (invitrogen)","Aβ plaque density by Thioflavin-S immunohistochemistry (stereology-based plaque count in visual cortex, hippocampus, PFC)","Tau phosphorylation at AT8 (Ser202/Thr205) and PHF-1 (Ser396/Ser404) epitopes by MSD electrochemiluminescence","Microglial morphology and phagocytosis: Iba1+ cell density, CD68/Iba1 ratio, TREM2 expression by qPCR and immunohistochemistry","PV+ interneuron count and firing properties (ex vivo patch clamp in acute brain slices)","LTP magnitude at Schaffer collateral-CA1 synapses (field EPSP slope, 100 Hz tetanus)","Novel object recognition (24-hour delay, discrimination index as %)","Morris water maze hidden platform (escape latency, probe trial % time in target quadrant)","40 Hz power spectral density in real-time EEG during stimulation (wireless telemetry)"],"expected_effect_size":"Cohen's d = 0.85 for hippocampal-cortical gamma coherence (primary), d = 1.0 for Aβ plaque reduction (literature-based, Martorell et al. 2019), d = 0.7 for cognitive improvement (NOR discrimination); power = 0.80, α = 0.05, accounting for multiple secondary outcomes via multiplicity correction","statistical_approach":"Mixed-effects two-way ANOVA (treatment × time) for longitudinal readouts; two-tailed Welch's t-test for post-hoc pairwise comparisons with Bonferroni-Dunn correction for 8 secondary outcomes (adjusted α = 0.00625); Kruskal-Wallis for non-normal distributions (plaque stereology); linear mixed models for LFP time-series with animal as random effect; all analyses pre-registered on Open Science Framework before data collection","sample_size_rationale":"Power calculation (G*Power 3.1): for two-group comparison with d=0.85, α=0.05, power=0.80, n=24/group minimum; inflated to n=30/group to accommodate mixed-effects model df loss and potential cage outliers; further inflated to n=35/group (15% buffer) given 6-week survival requirement; wild-type control n=20 sufficient for positive control verification (d=1.2 expected vs APP/PS1 baseline)","falsification_criterion":"Experiment falsifies hypothesis if: (1) hippocampal-prefrontal gamma PLV does not significantly differ between 40 Hz and sham groups (p > 0.00625 after multiplicity correction), OR (2) Aβ plaque burden shows no significant reduction in 40 Hz group vs sham (p > 0.00625), OR (3) novel object recognition discrimination index remains <55% in both groups (ceiling effect or no cognitive rescue); any single criterion met constitutes mechanistic failure and halts further dose-optimization experiments"},"created_at":"2026-04-28T13:00:29.816117-07:00","updated_at":"2026-04-28T13:00:29.816117-07:00","version_number":3,"parent_version_id":null,"version_tag":null,"changelog":null,"is_latest":1,"lifecycle_state":"active","superseded_by":null,"deprecated_at":null,"deprecated_reason":null,"dependencies":null,"market_price":0.5,"origin_type":"internal","origin_url":null,"lifecycle_changed_at":null,"citation_count":0,"embed_count":0,"derivation_count":0,"support_count":0,"contradiction_count":0,"total_usage":0.0,"usage_score":0.5,"usage_computed_at":null,"quality_status":null,"contributors":[],"answers_question_ids":null,"deprecated_reason_detail":null,"deprecated_reason_code":null,"commit_sha":null,"commit_submodule":null,"last_mutated_at":"2026-05-16T14:51:34.657673-07:00","disputed_at":null,"gap_id":null,"mission_id":null,"intrinsic_priority":null,"effective_priority":null,"artifact_id":"150212f7-a68f-4540-bff3-ae48ac6845eb","artifact_dir":null,"primary_filename":null,"accessory_filenames":null,"folder_layout_version":1,"migrated_to_folder_at":null,"hypothesis_id":null,"authorship":{"kind":"human","contributors":[{"role":"author","actor_ref":"forge_experiment_proposal_generator"}]},"epistemic_tier":"T3_provisional","created_by_agent_id":null},"outgoing_links":[{"target_artifact_id":"h-bdbd2120","link_type":"derives_from","strength":1.0,"evidence":"{\"source\": \"forge_experiment_proposal_generator\", \"method\": \"generate_experiment_proposal\"}"}],"incoming_links":[],"current_artifact_id":"experiment_proposal-afa252d5-e3ae-4839-9258-316fe0045c76","is_canonical":true,"supersede_chain":["experiment_proposal-afa252d5-e3ae-4839-9258-316fe0045c76"]}