{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T20:35:27Z","timestamp":1786739727637,"version":"build-2736575974"},"reference-count":45,"publisher":"Ovid Technologies (Wolters Kluwer Health)","issue":"10","license":[{"start":{"date-parts":[[2021,2,19]],"date-time":"2021-02-19T00:00:00Z","timestamp":1613692800000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":["lww.com","ovid.com"],"crossmark-restriction":true},"short-container-title":[],"abstract":"<jats:p>Inflammation and coagulation are tightly interconnected in the pathophysiology of neuronal diseases. Thrombin, a pro-coagulant serine protease is associated with neurodegeneration and its indirect inhibitor, activated protein C (aPC), is considered neuroprotective. While levels of thrombin and aPC activity are readily measured in the blood, similar assays in the cerebrospinal fluid (CSF) have not been described. The aim of this study was to establish a specific and sensitive enzymatic assay to measure both thrombin and aPC activity in the CSF. CSF was collected from 14 patients with suspected normal pressure hydrocephalus served as a control group, while seven patients with central nervous system infections served as an acute neuro-inflammatory study group and one sample of CSF following traumatic lumbar puncture served as a positive control. Thrombin and aPC activities were measured by fluorescence released by specific proteolytic cleavage in the presence of endopeptidase and amino-peptidase inhibitors to ensure specificity. Specificity of the method was verified by thrombin and serine-protease inhibitors N-alpha-((2-naphthylsulfinyl) glycyl)-DL-p-amidinophenylalanylpiperidine and phenylmethanesulfonyl fluoride. Inhibition of thrombin activity by CSF samples and levels of specific thrombin inhibitors were also assessed. Thrombin and aPC activities were reliably measured and were significantly higher in the CSF of patients with central nervous system infections compared to normal pressure hydrocephalus controls, suggesting the involvement of these factors in neuro-inflammation. CSF thrombin activity levels in the presence of known thrombin concentration were high in patients with central nervous system infections, and low in normal pressure hydrocephalus patients. Quantification of endogenous thrombin inhibitors protease nexin 1, amyloid precursor protein and anti-thrombin III in CSF by western blot indicated a significant elevation of amyloid precursor protein in infectious CSF. In conclusion, this study describes a novel and sensitive assay aimed at the detection of thrombin and aPC activity in CSF. This method may be useful for measuring these factors that reflect degenerative and protective influences of coagulation on neurological disorders. The study procedure was approved by the Ethics Committee of the Chaim Sheba Medical Center (approval No. 4245-17-SMC) on October 18, 2018.<\/jats:p>","DOI":"10.4103\/1673-5374.308098","type":"journal-article","created":{"date-parts":[[2021,2,18]],"date-time":"2021-02-18T15:09:31Z","timestamp":1613660971000},"page":"2086-2092","update-policy":"https:\/\/doi.org\/10.1097\/lww.0000000000001000","source":"Crossref","is-referenced-by-count":3,"title":["Markers for neural degeneration and regeneration: novel highly sensitive methods for the measurement of thrombin and activated protein C in human cerebrospinal fluid"],"prefix":"10.4103","volume":"16","author":[{"given":"Alexandra","family":"Gerasimov","sequence":"first","affiliation":[{"name":"Department of Neurology, The Chaim Sheba Medical Center, Ramat Gan, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Valery","family":"Golderman","sequence":"additional","affiliation":[{"name":"Department of Neurology and Neurosurgery, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shany Guly","family":"Gofrit","sequence":"additional","affiliation":[{"name":"Department of Neurology, The Chaim Sheba Medical Center, Ramat Gan, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shay Anat","family":"Aharoni","sequence":"additional","affiliation":[{"name":"Department of Neurology, The Chaim Sheba Medical Center, Ramat Gan, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniela Noa","family":"Zohar","sequence":"additional","affiliation":[{"name":"Department of Neurology, The Chaim Sheba Medical Center, Ramat Gan, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ze\u2019ev","family":"Itsekson-Hayosh","sequence":"additional","affiliation":[{"name":"Department of Neurology, The Chaim Sheba Medical Center, Ramat Gan, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tsviya","family":"Fay-Karmon","sequence":"additional","affiliation":[{"name":"Department of Neurology, The Chaim Sheba Medical Center, Ramat Gan, Israel"},{"name":"The Movement Disorders Institute, Chaim Sheba Medical Center, Ramat Gan, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sharon","family":"Hassin-Baer","sequence":"additional","affiliation":[{"name":"Department of Neurology, The Chaim Sheba Medical Center, Ramat Gan, Israel"},{"name":"Department of Neurology and Neurosurgery, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel"},{"name":"The Movement Disorders Institute, Chaim Sheba Medical Center, Ramat Gan, Israel"},{"name":"Sagol School of Neuroscience, Tel Aviv University, Tel Aviv, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joab","family":"Chapman","sequence":"additional","affiliation":[{"name":"Department of Neurology, The Chaim Sheba Medical Center, Ramat Gan, Israel"},{"name":"Department of Neurology and Neurosurgery, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel"},{"name":"Sagol School of Neuroscience, Tel Aviv University, Tel Aviv, Israel"},{"name":"Department of Physiology and Pharmacology, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel"},{"name":"Robert and Martha Harden Chair in Mental and Neurological Diseases, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicola","family":"Maggio","sequence":"additional","affiliation":[{"name":"Department of Neurology, The Chaim Sheba Medical Center, Ramat Gan, Israel"},{"name":"Department of Neurology and Neurosurgery, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel"},{"name":"Sagol School of Neuroscience, Tel Aviv University, Tel Aviv, Israel"},{"name":"Talpiot Medical Leadership Program, The Chaim Sheba Medical Center, Ramat Gan, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3373-8397","authenticated-orcid":false,"given":"Efrat","family":"Shavit-Stein","sequence":"additional","affiliation":[{"name":"Department of Neurology, The Chaim Sheba Medical Center, Ramat Gan, Israel"},{"name":"Department of Neurology and Neurosurgery, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"276","published-online":{"date-parts":[[2021,10]]},"reference":[{"key":"R1-20260526","doi-asserted-by":"crossref","first-page":"1277","DOI":"10.1212\/WNL.58.8.1277","article-title":"Increased levels of the pro-inflammatory prostaglandin PGE2 in CSF from ALS patients","volume":"58","author":"Almer","year":"2002","journal-title":"Neurology"},{"key":"R2-20260526","doi-asserted-by":"crossref","first-page":"643","DOI":"10.2174\/0929866525666180619101218","article-title":"Venom protein C activators as diagnostic agents for defects of protein C system","volume":"25","author":"Asmat","year":"2018","journal-title":"Protein Pept Lett"},{"key":"R3-20260526","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1080\/10408363.2017.1299682","article-title":"Insights into the human brain proteome: Disclosing the biological meaning of protein networks in cerebrospinal fluid","volume":"54","author":"Bastos","year":"2017","journal-title":"Crit Rev Clin Lab Sci"},{"key":"R4-20260526","doi-asserted-by":"crossref","first-page":"2347","DOI":"10.1097\/00001756-200108080-00013","article-title":"Quantitative measurements of mouse brain thrombin-like and thrombin inhibition activities","volume":"12","author":"Beilin","year":"2001","journal-title":"Neuroreport"},{"key":"R5-20260526","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1002\/jnr.20270","article-title":"Increased thrombin inhibition in experimental autoimmune encephalomyelitis","volume":"79","author":"Beilin","year":"2005","journal-title":"J Neurosci Res"},{"key":"R6-20260526","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1097\/WNR.0b013e328091c1e6","article-title":"Increased KPI containing amyloid precursor protein in experimental autoimmune encephalomyelitis brains","volume":"18","author":"Beilin","year":"2007","journal-title":"Neuroreport"},{"key":"R7-20260526","doi-asserted-by":"crossref","first-page":"844","DOI":"10.1007\/s12031-013-0072-y","article-title":"Quantitative detection of thrombin activity in an ischemic stroke model","volume":"51","author":"Bushi","year":"2013","journal-title":"J Mol Neurosci"},{"key":"R8-20260526","doi-asserted-by":"crossref","first-page":"138","DOI":"10.3389\/fneur.2017.00138","article-title":"A linear temporal increase in thrombin activity and loss of its receptor in mouse brain following ischemic stroke","volume":"8","author":"Bushi","year":"2017","journal-title":"Front Neurol"},{"key":"R9-20260526","doi-asserted-by":"crossref","first-page":"876","DOI":"10.1055\/s-0033-1357482","article-title":"Coagulation in inflammatory diseases of the central nervous system","volume":"39","author":"Chapman","year":"2013","journal-title":"Semin Thromb Hemost"},{"key":"R10-20260526","first-page":"527","article-title":"Progressive multiple sclerosis: from pathogenic mechanisms to treatment","volume":"140","author":"Correale","year":"2017","journal-title":"Brain"},{"key":"R11-20260526","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1002\/ana.24078","article-title":"Early detection of thrombin activity in neuroinflammatory disease","volume":"75","author":"Davalos","year":"2014","journal-title":"Ann Neurol"},{"key":"R12-20260526","doi-asserted-by":"crossref","first-page":"575","DOI":"10.1016\/0896-6273(91)90060-D","article-title":"Prothrombin mRNA is expressed by cells of the nervous system","volume":"6","author":"Dihanich","year":"1991","journal-title":"Neuron"},{"key":"R13-20260526","doi-asserted-by":"crossref","first-page":"26S","DOI":"10.1378\/chest.124.3_suppl.26S","article-title":"The protein C pathwaay","volume":"124","author":"Esmon","year":"2003","journal-title":"Chest"},{"key":"R14-20260526","doi-asserted-by":"crossref","first-page":"59","DOI":"10.3389\/fneur.2019.00059","article-title":"Thrombin and the coag-inflammatory nexus in neurotrauma ALS, and other neurodegenerative disorders","volume":"10","author":"Festoff","year":"2019","journal-title":"Front Neurol"},{"key":"R15-20260526","doi-asserted-by":"crossref","first-page":"e157","DOI":"10.1111\/j.1755-5949.2010.00183.x","article-title":"Neurosyphilis: A historical perspective and review","volume":"16","author":"Ghanem","year":"2010","journal-title":"CNS Neurosci Ther"},{"key":"R16-20260526","doi-asserted-by":"crossref","first-page":"2043","DOI":"10.4103\/1673-5374.262568","article-title":"The neuro-glial coagulonome: the thrombin receptor and coagulation pathways as major players in neurological diseases","volume":"14","author":"Gofrit","year":"2019","journal-title":"Neural Regen Res"},{"key":"R17-20260526","doi-asserted-by":"crossref","first-page":"2422","DOI":"10.3390\/ijms21072422","article-title":"A novel highly sensitive method for measuring inflammatory neural-derived APC activity in glial cell lines mouse brain and human CSF","volume":"21","author":"Golderman","year":"2020","journal-title":"Int J Mol Sci"},{"key":"R18-20260526","doi-asserted-by":"crossref","first-page":"1891","DOI":"10.1007\/s00604-016-1833-4","article-title":"Microplate based assay for thrombin detection using an RNA aptamer as affinity ligand and cleavage of a chromogenic or a fluorogenic peptide substrate","volume":"183","author":"Hao","year":"2016","journal-title":"Microchim Acta"},{"key":"R19-20260526","doi-asserted-by":"crossref","first-page":"1405","DOI":"10.4155\/bio-2019-0163","article-title":"Establishment of neurofilament light chain Simoa assay in cerebrospinal fluid and blood","volume":"11","author":"Hendricks","year":"2019","journal-title":"Bioanalysis"},{"key":"R20-20260526","doi-asserted-by":"crossref","first-page":"1385","DOI":"10.1212\/WNL.0b013e31828c2fda","article-title":"Idiopathic normal-pressure hydrocephalus Pathophysiology and diagnosis by CSF biomarkers","volume":"80","author":"Jeppsson","year":"2013","journal-title":"Neurology"},{"key":"R21-20260526","doi-asserted-by":"crossref","first-page":"1531","DOI":"10.1023\/A:1021147914843","article-title":"Prothrombin concentration in the cerebrospinal fluid is not altered in Alzheimer\u2019s disease","volume":"24","author":"Lewczuk","year":"1999","journal-title":"Neurochem Res"},{"key":"R22-20260526","doi-asserted-by":"crossref","first-page":"4113","DOI":"10.1021\/acs.jproteome.8b00300","article-title":"Deep Dive on the Proteome of Human Cerebrospinal Fluid: A Valuable Data Resource for Biomarker Discovery and Missing Protein Identification","volume":"17","author":"Macron","year":"2018","journal-title":"J Proteome Res"},{"key":"R23-20260526","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1001\/jama.2009.1064","article-title":"CSF biomarkers and incipient Alzheimer disease in patients with mild cognitive impairment","volume":"302","author":"Mattsson","year":"2009","journal-title":"JAMA"},{"key":"R24-20260526","doi-asserted-by":"crossref","first-page":"5237","DOI":"10.1182\/blood-2012-08-452169","article-title":"Biased agonism of protease-activated receptor 1 by activated protein C caused by noncanonical cleavage at Arg46","volume":"120","author":"Mosnier","year":"2012","journal-title":"Blood"},{"key":"R25-20260526","doi-asserted-by":"crossref","first-page":"6075","DOI":"10.1002\/anie.201007032","article-title":"Profiling of active thrombin in human blood by supramolecular complexes","volume":"50","author":"M\u00fcller","year":"2011","journal-title":"Angew Chem Int Ed Engl"},{"key":"R26-20260526","doi-asserted-by":"crossref","first-page":"2002","DOI":"10.1093\/brain\/awx120","article-title":"Brain inflammation accompanies amyloid in the majority of mild cognitive impairment cases due to Alzheimer\u2019s disease","volume":"140","author":"Parbo","year":"2017","journal-title":"Brain"},{"key":"R27-20260526","doi-asserted-by":"crossref","first-page":"e91974","DOI":"10.1371\/journal.pone.0091974","article-title":"Cerebrospinal fluid biomarker and brain biopsy findings in idiopathic normal pressure hydrocephalus","volume":"9","author":"Pyykk\u00f6","year":"2014","journal-title":"PLoS One"},{"key":"R28-20260526","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.brainres.2019.03.010","article-title":"Neuroprotection and vasculoprotection using genetically targeted protease-ligands","volume":"1715","author":"Rajput","year":"2019","journal-title":"Brain Res"},{"key":"R29-20260526","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1097\/00024382-200013020-00004","article-title":"Comparison of the mortality and inflammatory response of two models of sepsis: Lipopolysaccharide vs. cecal ligation and puncture","volume":"13","author":"Remick","year":"2000","journal-title":"Shock"},{"key":"R30-20260526","doi-asserted-by":"crossref","first-page":"510","DOI":"10.1007\/s00415-017-8730-6","article-title":"CSF neurofilament proteins as diagnostic and prognostic biomarkers for amyotrophic lateral sclerosis","volume":"265","author":"Rossi","year":"2018","journal-title":"J Neurol"},{"key":"R31-20260526","doi-asserted-by":"crossref","first-page":"1139","DOI":"10.3389\/fneur.2018.01139","article-title":"Blocking thrombin significantly ameliorates experimental autoimmune neuritis","volume":"9","author":"Shavit-Stein","year":"2019","journal-title":"Front Neurol"},{"key":"R32-20260526","doi-asserted-by":"crossref","first-page":"e0219453","DOI":"10.1371\/journal.pone.0219453","article-title":"The role of thrombin in the pathogenesis of diabetic neuropathy","volume":"14","author":"Shavit-Stein","year":"2019","journal-title":"PLoS One"},{"key":"R33-20260526","doi-asserted-by":"crossref","first-page":"3419","DOI":"10.3390\/ijms21103419","article-title":"Brain protease activated receptor 1 pathway: a therapeutic target in the superoxide dismutase 1 (SOD1) mouse model of amyotrophic lateral sclerosis","volume":"21","author":"Shavit-Stein","year":"2020","journal-title":"Int J Mol Sci"},{"key":"R34-20260526","first-page":"7692182","article-title":"Thrombin inhibition reduces the expression of brain inflammation markers upon systemic LPS treatment","author":"Shavit Stein","year":"2018","journal-title":"Neural Plast 2018"},{"key":"R35-20260526","doi-asserted-by":"crossref","first-page":"132","DOI":"10.3389\/fimmu.2012.00132","article-title":"Post-ischemic inflammation in the brain","volume":"3","author":"Shichita","year":"2012","journal-title":"Front Immunol"},{"key":"R36-20260526","doi-asserted-by":"crossref","first-page":"1163","DOI":"10.1182\/blood-2017-10-810895","article-title":"PAR1 biased signaling is required for activated protein C in vivo benefits in sepsis and stroke","volume":"131","author":"Sinha","year":"2018","journal-title":"Blood"},{"key":"R37-20260526","doi-asserted-by":"crossref","first-page":"1473","DOI":"10.1055\/s-0038-1665436","article-title":"Thrombin and its precursor in human cerebrospinal fluid","volume":"78","author":"Smirnova I","year":"1997","journal-title":"Thromb Haemost"},{"key":"R38-20260526","first-page":"564","article-title":"The comparison of selected cerebrospinal fluid and serum cytokine levels in patients with multiple sclerosis and normal pressure hydrocephalus","volume":"36","author":"Sosvorova","year":"2015","journal-title":"Neuroendocrinol Lett"},{"key":"R39-20260526","first-page":"586","article-title":"Selected pro- and anti-inflammatory cytokines in cerebrospinal fluid in normal pressure hydrocephalus","volume":"35","author":"Sosvorova","year":"2014","journal-title":"Neuroendocrinol Lett"},{"key":"R40-20260526","doi-asserted-by":"crossref","first-page":"324","DOI":"10.1177\/1076029606291381","article-title":"Specific determination of plasmatic thrombin activity","volume":"12","author":"Stief","year":"2006","journal-title":"Clin Appl Thromb"},{"key":"R41-20260526","doi-asserted-by":"crossref","first-page":"S96","DOI":"10.1016\/S0140-6736(17)30492-0","article-title":"Systemic inflammation and blood-brain barrier abnormality in relapsing-remitting multiple sclerosis","volume":"389","author":"Varatharaj","year":"2017","journal-title":"Lancet"},{"key":"R42-20260526","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.expneurol.2011.09.033","article-title":"Epilepsy and brain inflammation","volume":"244","author":"Vezzani","year":"2013","journal-title":"Exp Neurol"},{"key":"R43-20260526","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1093\/schbul\/sbx035","article-title":"Meta-analysis of cerebrospinal fluid cytokine and tryptophan catabolite alterations in psychiatric patients: comparisons between schizophrenia bipolar disorder, and depression","volume":"44","author":"Wang","year":"2018","journal-title":"Schizophr Bull"},{"key":"R44-20260526","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1016\/j.jinf.2006.11.016","article-title":"Procoagulant and fibrinolytic activity in cerebrospinal fluid from adults with bacterial meningitis","volume":"54","author":"Weisfelt","year":"2007","journal-title":"J Infect"},{"key":"R45-20260526","doi-asserted-by":"crossref","first-page":"577248","DOI":"10.1016\/j.jneuroim.2020.577248","article-title":"Anti-neurofilament antibodies and neurodegeneration: Markers and generators","volume":"344","author":"Zmira","year":"2020","journal-title":"J Neuroimmunol"}],"container-title":["Neural Regeneration Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.ovid.com\/10.4103\/1673-5374.308098","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T19:31:56Z","timestamp":1786735916000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.ovid.com\/10.4103\/1673-5374.308098"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,2,19]]},"references-count":45,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2021]]}},"URL":"https:\/\/doi.org\/10.4103\/1673-5374.308098","relation":{},"ISSN":["1673-5374","1876-7958"],"issn-type":[{"value":"1673-5374","type":"print"},{"value":"1876-7958","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,2,19]]},"assertion":[{"value":"2020-08-11","name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2020-11-12","name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}}]}}