Putative Biomarkers in Tears for Diabetic Retinopathy Diagnosis.

{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:35692536", "kind": "science-record", "revision": 5, "content_hash": "sha256:cf24b2d178b1f878d55d95140b149b35bcba8af311efe423dbda0ac63f5e95fc", "created_at": "2026-11-01T18:14:39Z", "updated_at": "2026-11-01T22:49:16Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "Putative Biomarkers in Tears for Diabetic Retinopathy Diagnosis.", "doi": "10.3389/fmed.2022.873483", "pmid": "35692536", "authors": ["Madania Amorim", "Beatriz Martins", "Francisco Caramelo", "Conceição Gonçalves", "Grimalde Trindade", "Jorge Simão", "Patrícia Barreto", "Inês Marques", "Ermelindo Carreira Leal", "Eugénia Carvalho", "Flávio Reis", "Teresa Ribeiro-Rodrigues", "Henrique Girão", "Paulo Rodrigues-Santos", "Cláudia Farinha", "António Francisco Ambrósio", "Rufino Silva", "Rosa Fernandes"], "journal": "Frontiers in medicine", "year": 2022, "abstract": "Purpose: Tear fluid biomarkers may offer a non-invasive strategy for detecting diabetic patients with increased risk of developing diabetic retinopathy (DR) or increased disease progression, thus helping both improving diagnostic accuracy and understanding the pathophysiology of the disease. Here, we assessed the tear fluid of nondiabetic individuals, diabetic patients with no DR, and diabetic patients with nonproliferative DR (NPDR) or with proliferative DR (PDR) to find putative biomarkers for the diagnosis and staging of DR.\nMethods: Tear fluid samples were collected using Schirmer test strips from a cohort with 12 controls and 54 Type 2 Diabetes (T2D) patients, and then analyzed using mass spectrometry (MS)-based shotgun proteomics and bead-based multiplex assay. Tear fluid-derived small extracellular vesicles (EVs) were analyzed by transmission electron microscopy, Western Blotting, and nano tracking.\nResults: Proteomics analysis revealed that among the 682 reliably quantified proteins in tear fluid, 42 and 26 were differentially expressed in NPDR and PDR, respectively, comparing to the control group. Data are available via ProteomeXchange with identifier PXD033101. By multicomparison analyses, we also found significant changes in 32 proteins. Gene ontology (GO) annotations showed that most of these proteins are associated with oxidative stress and small EVs. Indeed, we also found that tear fluid is particularly enriched in small EVs. T2D patients with NPDR have higher IL-2/-5/-18, TNF, MMP-2/-3/-9 concentrations than the controls. In the PDR group, IL-5/-18 and MMP-3/-9 concentrations were significantly higher, whereas IL-13 was lower, compared to the controls.\nConclusions: Overall, the results show alterations in tear fluid proteins profile in diabetic patients with retinopathy. Promising candidate biomarkers identified need to be validated in a large sample cohort.", "claims": [{"id": "claim-method", "statement": "A t-test was performed (FDR = 0.05 and s0 = 1) to compare tear samples between control group and T2D, NPDR and PDR groups, and between NDPR and PDR groups, and a volcano plot was generated. For normal distribution, one-way ANOVA followed by post-hoc analysis (Tukey test) was carried out to test for significance for a specific protein or analyte. Whenever a variable did not reach the normality assumption the non-parametric Kruskal-Wallis test followed by Dunn-Sidak test post-hoc test was performed. The diagnostic power of biomarkers was evaluated with receiver operating characteristics (ROC) curves (AUC, confidence interval).", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:35692536", "locator": "sections[16] \"Statistical Analysis\" paragraph 2", "quote": "A t-test was performed (FDR = 0.05 and s0 = 1) to compare tear samples between control group and T2D, NPDR and PDR groups, and between NDPR and PDR groups, and a volcano plot was generated."}, {"asset": "source:pmid:35692536", "locator": "sections[16] \"Statistical Analysis\" paragraph 2", "quote": "For normal distribution, one-way ANOVA followed by post-hoc analysis (Tukey test) was carried out to test for significance for a specific protein or analyte."}, {"asset": "source:pmid:35692536", "locator": "sections[16] \"Statistical Analysis\" paragraph 2", "quote": "Whenever a variable did not reach the normality assumption the non-parametric Kruskal-Wallis test followed by Dunn-Sidak test post-hoc test was performed."}, {"asset": "source:pmid:35692536", "locator": "sections[16] \"Statistical Analysis\" paragraph 2", "quote": "The diagnostic power of biomarkers was evaluated with receiver operating characteristics (ROC) curves (AUC, confidence interval)."}], "state": "complete"}, {"id": "claim-data-access", "statement": "The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE (82) partner repository with the dataset identifier PXD033101 and 10.6019/PXD033101.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:35692536", "locator": "sections[23] \"Data Availability Statement\" paragraph 2", "quote": "The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE (82) partner repository with the dataset identifier PXD033101 and 10.6019/PXD033101."}], "state": "complete"}, {"id": "claim-target-result", "statement": "Multicomparison analysis identified significant changes in 32 tear-fluid proteins.", "kind": "qualitative", "target": null, "evidence": [{"asset": "source:pmid:35692536", "locator": "sections[0] \"Abstract\" paragraph 3", "quote": "By multicomparison analyses, we also found significant changes in 32 proteins."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "partial", "objective": "Multicomparison analysis identified significant changes in 32 tear-fluid proteins.", "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:35692536"], "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. This lane did not download the dataset or read its licence.", "duration_hours": null, "parameters": {}, "inputs": ["data:geo:PXD033101"], "outputs": ["the source data with its repository provenance"], "instruments": [], "reagents": []}, {"id": "run_analysis", "order": 3, "name": "Run the published analysis", "kind": "analysis", "state": "blocked", "blocked_on": "method:student_t", "description": "Apply the paper's statistical method to the resolved series and compare the result with the published target.", "duration_hours": null, "parameters": {}, "inputs": ["data:geo:PXD033101"], "outputs": ["the replicated result and its comparison"], "instruments": [], "reagents": []}], "materials": [], "instruments": [], "reagents": [], "conditions": {}, "gaps": ["step:run_analysis blocked on method:student_t"]}, "data": {"state": "partial", "assets": [{"id": "source:pmid:35692536", "role": "manuscript", "format": "xml", "media_type": "application/xml", "uri": "https://doi.org/10.3389/fmed.2022.873483", "sha256": "sha256:1d467b422c5af4483caef5aeb6ec5fa9399769f55665ee50c3c859dea0de5bc4", "bytes": 80734, "access": "open", "state": "resolved", "reader": null, "reader_status": "reader_required", "identifier": "10.3389/fmed.2022.873483", "version": null, "note": "Full article fetched from PMC; the checksum covers the rendered section body, not the original XML bytes.", "structure": {"pmcid": "PMC9174990", "source": "europe_pmc_fulltext", "coverage": "whole_body", "text_hash_scope": "rendered_section_body"}}, {"id": "data:geo:PXD033101", "role": "raw", "format": "geo-series", "media_type": "application/vnd.ncbi.geo-series+json", "uri": "https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=PXD033101", "sha256": null, "bytes": null, "access": "open", "state": "resolved", "reader": null, "reader_status": "reader_required", "identifier": "PXD033101", "version": null, "note": "The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE (82) partner repository with the dataset identifier PXD033101 and 10.6019/PXD033101.", "structure": {"repository": "GEO", "access_route": "GEO accession URL", "access_statement_verbatim": "The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE (82) partner repository with the dataset identifier PXD033101 and 10.6019/PXD033101.", "access_statement_kind": "data_availability_statement", "sample_accessions": [], "sample_count": null, "group_sizes": null, "group_sizes_basis": null, "platform": null, "taxon": null, "title": "Putative tear fluid biomarkers of diabetic retinopathy", "resolution_checked_at": "2026-11-01T18:06:51Z", "resolution_check": "identifier_resolution_only_no_download_no_licence_read", "downloaded": false, "licence_read": false}}], "gaps": ["asset:data:geo:PXD033101 not downloaded", "asset:data:geo:PXD033101 licence not read"]}, "analysis": {"state": "partial", "methods": [{"id": "method-1", "family": "group_comparison", "specific": "student_t_test", "label": "Group comparison", "dsl_operations": [], "missing_operations": ["student_t"], "expressible": false, "parameters": {}, "data_shapes": ["two_or_more_groups_continuous"], "evidence": {"score": 1.0, "band": "reordered", "quote": "A t-test was performed (FDR = 0.05 and s0 = 1) to compare tear samples between control group and T2D, NPDR and PDR groups, and between NDPR and PDR groups, and a volcano plot was generated. For normal distribution, one-way ANOVA followed by post-hoc analysis (Tukey test) was carried out to test for significance for a specific protein or analyte. Whenever a variable did not reach the normality assumption the non-parametric Kruskal-Wallis test followed by Dunn-Sidak test post-hoc test was performed. 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Tear fluid-d", "changes in tear samples were evaluated using mass spectrometry (MS)-based shotgun proteomics. A total of 32", "mics. A total of 32 samples was prepared for LC-MS/MS analyses. Proteins were denatured by additio"], "order_basis": "paper_text_order:718", "id": "assay:1"}, {"role": "data_generation", "definition_id": "cell_imaging", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.69, "evidence": ["(EVs) were analyzed by transmission electron microscopy, Western Blotting, and nano tracking. Result"], "order_basis": "paper_text_order:891", "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": 0.93, "evidence": ["nalyzed by transmission electron microscopy, Western Blotting, and nano tracking. Results: Proteomics anal"], "order_basis": "paper_text_order:903", "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": ["A t-test was performed (FDR = 0.05 and s0 = 1) to compare tear samples between control group and T2D, NPDR and PDR groups, and between NDPR and PDR groups, and a volcano plot was generated. 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