{"data":{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:32886703", "kind": "science-record", "revision": 7, "content_hash": "sha256:c9436a070d9a430fdb179472f5788185ad5b02a3cca83d3d2e7ccc0f0b1d4046", "created_at": "2026-11-01T15:41:09Z", "updated_at": "2026-11-01T22:02:08Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "New label-free methods for protein relative quantification applied to the investigation of an animal model of Huntington Disease.", "doi": "10.1371/journal.pone.0238037", "pmid": "32886703", "authors": ["Flora Cozzolino", "Alfredo Landolfi", "Ilaria Iacobucci", "Vittoria Monaco", "Marianna Caterino", "Simona Celentano", "Chiara Zuccato", "Elena Cattaneo", "Maria Monti"], "journal": "PloS one", "year": 2020, "abstract": "Spectral Counts approaches (SpCs) are largely employed for the comparison of protein expression profiles in label-free (LF) differential proteomics applications. Similarly, to other comparative methods, also SpCs based approaches require a normalization procedure before Fold Changes (FC) calculation. Here, we propose new Complexity Based Normalization (CBN) methods that introduced a variable adjustment factor (f), related to the complexity of the sample, both in terms of total number of identified proteins (CBN(P)) and as total number of spectral counts (CBN(S)). Both these new methods were compared with the Normalized Spectral Abundance Factor (NSAF) and the Spectral Counts log Ratio (Rsc), by using standard protein mixtures. Finally, to test the robustness and the effectiveness of the CBNs methods, they were employed for the comparative analysis of cortical protein extract from zQ175 mouse brains, model of Huntington Disease (HD), and control animals (raw data available via ProteomeXchange with identifier PXD017471). LF data were also validated by western blot and MRM based experiments. On standard mixtures, both CBN methods showed an excellent behavior in terms of reproducibility and coefficients of variation (CVs) in comparison to the other SpCs approaches. Overall, the CBN(P) method was demonstrated to be the most reliable and sensitive in detecting small differences in protein amounts when applied to biological samples.", "claims": [{"id": "claim-method", "statement": "Mass spectrometry data (LF) statistical significance was determined by the unpaired Student’s t-test and differences were considered statistically significant at Benjamini-Hochberg corrected p-value (FDR)<0.05 [44].", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:32886703", "locator": "sections[11] \"Statistical analyses\" paragraph 2", "quote": "Mass spectrometry data (LF) statistical significance was determined by the unpaired Student’s t-test and differences were considered statistically significant at Benjamini-Hochberg corrected p-value (FDR)<0.05 [44]."}], "state": "complete"}, {"id": "claim-data-access", "statement": "Data are available via ProteomeXchange with identifier PXD017471.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:32886703", "locator": "sections[21] \"Data Availability\" paragraph 2", "quote": "Data are available via ProteomeXchange with identifier PXD017471."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "partial", "objective": null, "steps": [{"id": "read_published_estimate", "order": 1, "name": "Read the published estimate from the source", "kind": "other", "state": "complete", "blocked_on": null, "description": "The claim, method and evidence span published with the source article.", "duration_hours": null, "parameters": {}, "inputs": ["source:pmid:32886703"], "outputs": ["the published estimate and its evidence"], "instruments": [], "reagents": []}, {"id": "obtain_source_data", "order": 2, "name": "Obtain the source data from the named repository", "kind": "data-preparation", "state": "complete", "blocked_on": null, "description": "The GEO series accession resolves and its sample accessions are listed. 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"Application of CBN methods to the differential proteomics analysis of samples from cerebral cortex from Huntington’s Disease mice", "row_count": 14, "column_count": 7}, "table": {"label": "Table 2", "caption": "Summary of proteins FCs validated by western blot and/or MRM.", "rows": [["Protein", "FC MRM", "FC WB", "FC CBN(S)", "FC CBN(P)", "FC RSC", "FC NSAF"], ["HTT", "0.59", "0.42", "0.08", "0.59", "0.14", "/"], ["CAMK2A", "0.44", "/", "/", "0.67", "/", "/"], ["SAMM50", "0.80", "0.66", "0.44", "0.69", "0.47", "/"], ["ATP2A2", "0.69", "/", "/", "0.69", "/", "/"], ["HOMER1", "0.60", "0.72", "0.45", "0.71", "0.45", "/"], ["IRGM1", "/", "1.93", "19.73", "1.22", "4.09", "UP"], ["APO-A4", "1.27", "/", "/", "1.29", "/", "/"], ["RIC8A", "1.30", "/", "4.46", "1.30", "/", "/"], ["STIP1", "1.35", "/", "2.38", "1.32", "2.11", "2.50"], ["OSBPL2", "1.32", "1.18", "6.37", "/", "2.92", "/"], ["SERPIN B6", "1.33", "1.99", "2.55", "/", "2.02", "2.75"], ["HNRNP H", "/", "0.46", "0.30", "/", "/", "/"], ["UQCRQ", "/", "0.71", "0.36", "/", "/", "/"]], "row_count": 14, "column_count": 7, "numeric_cells": 47, "header_rows": 1, "locator": "section[17]/table-wrap[1]#pone.0238037.t002"}}, {"id": "asset:table:pmid:32886703:3", "role": "table", "format": "jats-table", "media_type": "application/vnd.jats.table+json", "uri": null, "sha256": "sha256:7b054b570ab397ad23e377520f454af3e91fc97e479b2bbe52255c6d4f071575", "bytes": 1157, "access": "open", "state": "verified", "reader": "read_table", "reader_status": "available", "identifier": "section[17]/table-wrap[2]#pone.0238037.t003", "version": null, "note": "List of functional pathways identified by STRING analysis.", "structure": {"pmcid": "PMC7473538", "section_index": 17, "section_title": "Application of CBN methods to the differential proteomics analysis of samples from cerebral cortex from Huntington’s Disease mice", "row_count": 10, "column_count": 9}, "table": {"label": "Table 3", "caption": "List of functional pathways identified by STRING analysis.", "rows": [["Pathways Description", "Observed Gene Count (NSAF)", "FDR", "Observed Gene Count (RSC)", "FDR", "Observed Gene Count (CBN(P))", "FDR", "Observed Gene Count (CBN(S))", "FDR"], ["Metabolic pathways", "19", "2.62E-09", "42", "1.10E-09", "22", "2.30E-06", "42", "1.51E-10"], ["Oxidative phosphorylation", "7", "0.0037", "14", "9.68E-09", "7", "0.000105", "12", "7.09E-07"], ["Parkinson's disease", "9", "0.00049", "14", "1.77E-08", "8", "1.53E-05", "12", "1.11E-06"], ["Huntington's disease", "9", "0.00156", "14", "2.78E-07", "9", "9.82E-06", "13", "1.24E-06"], ["Alzheimer's disease", "9", "0.00111", "13", "8.40E-07", "7", "0.000397", "12", "3.94E-06"], ["Calcium signaling pathway", "8", "0.00412", "7", "0.0328", "7", "0.000566", "8", "0.0109"], ["Val, Leu and Ile degradation", "6", "0.00049", "5", "0.00648", "3", "0.0269", "5", "0.00578"], ["cGMP-PKG signaling pathway", "8", "0.0037", "8", "0.0094", "5", "0.02", "7", "0.0247"], ["Fatty acid degradation", "5", "0.00194", 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"missing_operations": [], "expressible": true, "parameters": {}, "data_shapes": ["two_or_more_groups_continuous"], "evidence": {"score": 0.8, "band": "high", "quote": "Welch"}}, {"id": "method-3", "family": "group_comparison", "specific": "mann_whitney", "label": "Group comparison", "dsl_operations": [], "missing_operations": ["group_comparison_mann_whitney"], "expressible": false, "parameters": {}, "data_shapes": ["two_or_more_groups_continuous"], "evidence": {"score": 0.8, "band": "high", "quote": "Mann-Whitney"}}, {"id": "method-4", "family": "regression_modelling", "specific": "ols", "label": "Regression modelling", "dsl_operations": ["ols_fit"], "missing_operations": [], "expressible": true, "parameters": {}, "data_shapes": ["predictors_and_outcome"], "evidence": {"score": 0.8, "band": "high", "quote": "Linear regression"}}, {"id": "method-5", "family": "image_quantification", "specific": "densitometry", "label": "Image analysis / quantification", "dsl_operations": 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the method it used and the corpus's method extractor maps it onto the DSL catalogue: student_t_test."}, "capability": {"tier": "R1", "capability_id": "capability:computational_analysis", "availability_state": "owned", "basis": "The computational analysis is owned. The physical bench the paper used is not, so a physical validation cannot be run here."}, "data": {"tier": "R1", "reachable": true, "accessions": ["PXD017471"], "resolved_accessions": ["PXD017471"], "statements": ["Data are available via ProteomeXchange with identifier PXD017471."], "basis": "The paper names PXD017471 in ProteomeXchange and the accession resolves in the repository. No dataset was downloaded and no licence was read."}}, "can_run": {"dsl_operations": [], "assets": ["source:pmid:32886703", "data:geo:PXD017471", "asset:table:pmid:32886703:1", "asset:table:pmid:32886703:2", "asset:table:pmid:32886703:3"], "steps": []}, "must_build": {"dsl_operations": [{"operation": "student_t", "unlocks_papers": null, "priority": null}], "converters": [], "readers": [], "capabilities": [], "data_gaps": ["data:geo:PXD017471"]}, "gaps": []}, "review": {"state": "partial", "machine_critique": {"kind": "extraction_review", "reader": {"model": "MiniMax-M3.1-Flash", "provider": "minimax", "provider_family": "minimax", "read_at": "2026-10-31T18:22:15Z", "independence": "single_reader"}, "evidence_check": {"method": "verbatim", "availability": "verbatim"}, "calls": 3, "usage": {"prompt_tokens": 49718, "completion_tokens": 961, "cached_tokens": 16648}}, "human_critique": null, "stages": [{"id": "extraction", "state": "complete", "reader": "MiniMax-M3.1-Flash", "provider": "minimax"}], "gaps": ["review.independent_reader", "review.machine_critique", "review.human_critique"]}, "execution": {"state": "absent", "status": "not_run", "mode": null, "runs": [], "gaps": ["execution.not_run"]}, "provenance": {"entities": [{"id": "pmid:32886703", "type": "paper", "label": "pmid:32886703"}, {"id": "source:pmid:32886703", "type": "manuscript", "label": "PMID 32886703 full text"}, {"id": "data:geo:PXD017471", "type": "dataset", "label": "GEO PXD017471"}], "activities": [{"id": "activity-reading-sci-2026-11-01", "type": "classification", "used": [{"id": "pmid:32886703", "type": "paper", "label": "pmid:32886703"}, {"id": "source:pmid:32886703", "type": "manuscript", "label": "PMID 32886703 full text"}, {"id": "data:geo:PXD017471", "type": "dataset", "label": "GEO PXD017471"}], "agent": "agent:corpus-assay-engine"}, {"id": "activity-read-32886703", "type": "extraction", "used": ["source:pmid:32886703"], "agent": "agent:minimax-reader"}, {"id": "activity-resolve-PXD017471", "type": "accession_resolution", "used": ["data:geo:PXD017471"], "agent": "agent:corpus-assay-engine"}], "agents": [{"id": "agent:corpus-assay-engine", "type": "software", "label": "Corpus assay engine"}, {"id": "agent:minimax-reader", "type": "software", "label": "MiniMax literature reader"}], "source_run_id": "reading-sci-2026-11-01", "dsl_catalogue_fingerprint": "343fb30756ba6ec3", "capability_table_fingerprint": "2cb3e42f047d2ecf", "snapshot_id": "ba34b49490d9bfaf"}, "lineage": {"parent_id": "pmid:32886703", "parent_hash": "sha256:b3ddae108fb52cc939d74e071b5828bc9b8ce0e1c4505b02fb61bf34276a5738", "fork_of": null, "generation": 6}, "gaps": ["asset:data:geo:PXD017471 licence not read", "asset:data:geo:PXD017471 not downloaded", "execution.not_run", "method.student_t", "review.human_critique", "review.independent_reader", "review.machine_critique", "step:run_analysis blocked on method:student_t"], "assays": [{"role": "data_generation", "definition_id": "mass_spectrometry", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["ion profiles in label-free (LF) differential proteomics applications. Similarly, to other comparativ", "tion of Spectral Counts data in differential proteomics experiments to calculate the relative Fold C", "isting normalization methods in differential proteomics experiments using the label-free procedure a", "plication of CBN methods to the differential proteomics analysis of samples from cerebral cortex fro"], "order_basis": "paper_text_order:222", "id": "assay:1"}, {"role": "data_generation", "definition_id": "animal_model", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["lysis of cortical protein extract from zQ175 mouse brains, model of Huntington Disease (HD), an", "d proteins with all methods in the WT and HD mouse model.(A) The histogram shows for each method the", "whose expression was instead decreased in HD mice were evaluated on total protein extracts of three", "oteins differentially expressed in WT and HD mouse model.(A) Western blot assays performed on total p"], "order_basis": "paper_text_order:984", "id": "assay:2"}, {"role": "data_generation", "definition_id": "western_blot", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["r PXD017471). LF data were also validated by western blot and MRM based experiments. On standard mixtu", "teins by two independent methodologies, i.e. western blot analyses and Multiple Reaction Monitoring (M", "itoring (MRM) mass spectrometry analysis. In western blot assays (Fig 5A), the expression levels of IR", "ially expressed in WT and HD mouse model.(A) Western blot assays performed on total protein extracts f"], "order_basis": "paper_text_order:1151", "id": "assay:3"}, {"role": "analysis", "definition_id": "group_comparison", "definition_version": "f2132e96ae6c7a4e", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"dsl_catalogue": "f2132e96ae6c7a4e"}, "confidence": 1.0, "evidence": ["Mass spectrometry data (LF) statistical significance was determined by the unpaired Student’s t-test and differences were considered statistically significant at Benjamini-Hochberg corrected p-value (FDR)<0.05 [44]."], "order_basis": "consumes_earlier_output", "id": "assay:4"}]},"meta":{"api_version":"v1","automation_ladder_snapshot":"ba34b49490d9bfaf","automation_ladder_caveats":["Method extraction is rule-based and has no hand-judged accuracy sample. Evidence bands order textual support; method agreement intervals are null (n=0), never probabilities. Work-type validation does not validate methods.","Operation coverage describes only explicitly named methods. It does not prove a complete protocol, compatible parameters, successful compilation or re-execution. Unrecognised or ambiguous methods require review.","Data reachability is a textual accession or public-data statement. No dataset was downloaded or checked for access, completeness or licence.","Capabilities are planning states seeded from the dated equipment register. No bench ownership, completed quote or partner booking is inferred. Procurement counts are dependencies, not guaranteed replications or financial returns.","Tier and blocker counts are deterministic assessments of this corpus, not measured agreement. R3 is reserved for procurement; unresolved methods and data require human work (R4). The most restrictive axis wins, with capability, method, data as tie-breaks. Resolving one blocker can leave another at the same tier."],"engine":"corpus_engine","engine_provider":"rule_based_v1+lex99e1195c7d6f","run_id":"run-20261023T122449232913","source":"esearch","source_detail":"PubMed mirror biomed_pubmed (read-only)","run_status":"complete","imported_at":"2026-11-03T20:16:52+00:00","generated_at":"2026-11-04T01:28:21+00:00","total_papers":5529,"total_assays_tracked":29,"automatable_assay_families":6,"validated":true,"caveat_count":9,"blocking_caveats":5,"caveats":[{"code":"low_evidence_dominates","basis":["4,152 of 5,529 classifications (75.1%) matched no cue evidence and sit at the 0.05 floor.","34 of 230 data_only calls (14.8%) are in that group."],"claim":"Work type and confidence bands","gloss":"Most records were classified from their title and framing alone, with assay-cue coverage at zero.","detail":"Most classifications in this run rest on no matched evidence at all. At the evidence floor the work type comes from title framing and a fixed prior, not from anything in the paper, so the published work-type split should be read as a rough triage signal rather than a measured distribution. The impact is bounded but not removed: since 2026-10-23 the engine excludes floor-confidence records from the shortlist entirely and publishes them separately as held out, so the recommendation is unaffected even though the census is.","status":"unverified","severity":"high"},{"code":"runtime_is_modelled","basis":["Runtime is summed from published per-assay hour figures. Timing awaits the first executed replication.","4,407 of 5,529 records (79.7%) model to 0.00 hours because no assay was matched, which the ranking then reads as \"cheapest\".","0 of 10 shortlisted records carry a 0.00-hour estimate."],"claim":"Estimated replication runtime","gloss":"The hour figures are estimates built from assay mentions; timing awaits the first executed replication.","detail":"The hour figures are a cost model derived from the bench steps implied by each matched assay family. The first executed replication will supply measured timing. Records without an assay match currently receive a zero-hour model value.","status":"unverified","severity":"high"},{"code":"signal_score_without_citations","basis":["5,529 of 5,529 records (100.0%) have no citation count from the source metadata."],"claim":"Signal score","gloss":"The signal score uses publication-type weight because the mirror supplied zero citation counts.","detail":"The source supplied no citation counts for any record in this run, so the signal score is publication-type weight only. The citation term of the ranking is inert here, and the shortlist is ordered by publication type alone.","status":"unverified","severity":"high"},{"code":"abstract_absent","basis":["1,667 of 5,529 records (30.2%) carry no abstract at all."],"claim":"Evidence base for classification","gloss":"Some records contain a title and publication type only, defining the available classification evidence.","detail":"Where the mirror contains a title and publication types only, those fields define the classifier evidence. A methods section is required for fuller automated and human review.","status":"unverified","severity":"medium"},{"code":"classifier_agreement_measured","basis":["Published two-way decision agreement 77.7% (310 of 399 decidable records) from a stratified hand-check of 438 real records.","Three-way diagnostic agreement 73.9% (288 of 390 decidable records).","data_only agreement 75.2% (100 of 133 decidable).","48 record(s) could not be decided from title and abstract and were excluded from the denominator.","56 analysis-only records were published under another work type, so the data_only share is not an upper bound."],"claim":"Published automatable decision","gloss":"This figure has been checked against a human reading the same papers, so we know how often it is right.","detail":"The published automatable / not-automatable distribution comes from a rule-based classifier that was hand-checked against a fixed ground-truth definition. The agreement rates below are measured, not estimated. The three-way work type remains a diagnostic and is reported separately.","status":"measured","severity":"high"},{"code":"llm_comparison_not_measured","basis":["rule_based_v1+lex99e1195c7d6f produced 5,529 of 5,529 classifications.","Measured on 438 labelled records; these are RULE-BASED figures, not model results."],"claim":"Rule-based versus LLM agreement","gloss":"Every number here is the rule engine’s own. Model comparison begins when a working API credential is connected and measured.","detail":"The published two-way decision agreed with a human reading on 310 of 399 decidable records (77.7%). The diagnostic three-way label agreed on 288 of 390 decidable records (73.9%). Every percentage in this run belongs to the rule engine. Model comparison begins when a working API credential is connected and measured.","status":"not_measured","severity":"high"},{"code":"confidence_is_a_band","basis":["low band: 135 of 210 hand-judged records were correct, 64.3% [57.6, 70.5] (95% CI).","medium band: 80 of 104 hand-judged records were correct, 76.9% [68.0, 84.0] (95% CI).","high band: 73 of 76 hand-judged records were correct, 96.0% [89.0, 98.7] (95% CI).","A fitted calibrated probability was built, measured and withdrawn: it overstated held-out accuracy and reorders nothing."],"claim":"Confidence band and evidence score","gloss":"The confidence band carries a measured accuracy interval; 0.85 represents relative rank strength.","detail":"Confidence on this run is a BAND plus its measured accuracy interval. The number stored alongside each record is a monotone evidence score that orders records; 0.85 represents relative rank strength. A fitted probability was withdrawn after it overstated held-out accuracy. Threshold on the band.","status":"measured","severity":"medium"},{"code":"three_way_split_is_diagnostic","basis":["The corpus is published at the two-way decision: automatable or not-automatable. The three-way split (data_only / physical / mixed) is published as a diagnostic with its measured reliability attached, not as a claim, because the rule engine's `mixed` call cannot be defended.","The engine called 22 record(s) mixed; 10 matched the human judgement (45.5%, 95% CI 26.9-65.3).","The engine's `mixed` call is right on 10/22 hand-judged records (45.5%, 95% CI 26.9%-65.3%); a constant `physical` guess scores 36.4%.","The engine's `mixed` call does not reliably beat a constant `physical` guess on the hand-judged records it labels mixed.","The sign of that difference flips between two independent samples, so the three-way refinement is not demonstrably informative.","The mixed stratum contributes a disproportionate share of errors relative to its size.","At title-only depth the physical/mixed split collapses: a title says that work was done, not whether it was also analysed."],"claim":"The published decision is two-way","gloss":"The three-way split is shown for diagnosis only. The published split is two ways.","detail":"Count and filter the corpus on automatable versus not-automatable. The data_only / physical / mixed split remains an internal engine diagnostic because the mixed call has lower measured reliability.","status":"measured","severity":"medium"},{"code":"automatable_flag_unproven","basis":["6 of 29 assay families are marked automatable now: meta_analysis, machine_learning, survival_analysis, pca, molecular_docking, differential_expression.","The flag is a rule over assay family and work type; validation begins with the first executed replication.","It inherits the 75.2% data_only agreement rate, because the flag requires the record to be analysis-only."],"claim":"Automatable now","gloss":"This flag marks assay families the rules classify as data-only. The first executed replication will validate it.","detail":"This flag marks families the rules consider replicable from data alone. Its validity depends on the analysis-only classification and will be measured through an executed replication.","status":"unverified","severity":"medium"}],"confidence_policy":"band_with_measured_interval","confidence_contract":{"policy":"band_with_measured_interval","summary":"Confidence is reported as a BAND with a measured accuracy interval. The raw evidence score supplies monotone ordering. A fitted point estimate was withdrawn after it overstated held-out accuracy.","evidence_score":"A monotone evidence score in [0,1] built from cue mass, axis separation and text coverage. Use it to order records. Interpret 0.85 as relative rank strength.","confidence_band":"low | medium | high. Consumers threshold on this value.","band_accuracy":"Wilson 95% interval on the measured agreement of the hand-judged records in this band.","calibrated_confidence":"WITHDRAWN. Reintroduction requires held-out evidence demonstrating reliable estimates across every bin."},"band_accuracy":{"low":{"n":210,"low":0.576,"band":"low","high":0.7046,"agree":135,"accuracy":0.6429},"high":{"n":76,"low":0.8903,"band":"high","high":0.9865,"agree":73,"accuracy":0.9605},"medium":{"n":104,"low":0.6795,"band":"medium","high":0.8397,"agree":80,"accuracy":0.7692}},"provider":"rule_based_v1+lex99e1195c7d6f","lexicon_id":"99e1195c7d6f","contract_published_at":"2026-10-23T12:25:29.022232+00:00","confidence_is_band":true,"confidence_is_probability":false,"decision_taxonomy":"two_way","decidable_fraction_two_way":0.8854,"decidable_fraction_two_way_low":0.8565,"decidable_fraction_two_way_high":0.9143,"decidable_fraction_three_way":0.8586,"decidable_fraction_three_way_low":0.8287,"decidable_fraction_three_way_high":0.8884,"decidable_population":5529,"validation_sample_size":438,"three_way_cost_fraction":0.0268,"three_way_cost_records":148,"decision_counts":{"automatable":230,"not_automatable":5299},"decision_split":{"automatable":{"count":230,"share":0.04159884246699222},"not_automatable":{"count":5299,"share":0.9584011575330078}},"diagnostic_work_type_counts":{"mixed":24,"physical":5275,"data_only":230}}}