{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,18]],"date-time":"2026-04-18T04:07:08Z","timestamp":1776485228009,"version":"3.51.2"},"reference-count":40,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2020,3,31]],"date-time":"2020-03-31T00:00:00Z","timestamp":1585612800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Background: Neural inflammation is linked to coagulation. Low levels of thrombin have a neuroprotective effect, mediated by activated protein C (APC). We describe a sensitive novel method for the measurement of APC activity at the low concentrations found in neural tissue. Methods: APC activity was measured using a fluorogenic substrate, Pyr-Pro-Arg-AMC, cleaved preferentially by APC. Selectivity was assessed using specific inhibitors and activators. APC levels were measured in human plasma, in glia cell lines, in mice brain slices following mild traumatic brain injury (mTBI) and systemic lipopolysaccharide (LPS) injection, and in cerebrospinal fluid (CSF) taken from viral meningoencephalitis patients and controls. Results: Selectivity required apixaban and alpha-naphthylsulphonylglycyl-4-amidinophenylalanine piperidine (NAPAP). APC levels were easily measurable in plasma and were significantly increased by Protac and CaCl2. APC activity was significantly higher in the microglial compared to astrocytic cell line and specifically lowered by LPS. Brain APC levels were higher in posterior regions and increased by mTBI and LPS. Highly elevated APC activity was measured in viral meningoencephalitis patients CSF. Conclusions: This method is selective and sensitive for the measurement of APC activity that significantly changes during inflammation in cell lines, animal models and human CSF.<\/jats:p>","DOI":"10.3390\/ijms21072422","type":"journal-article","created":{"date-parts":[[2020,4,1]],"date-time":"2020-04-01T03:44:13Z","timestamp":1585712653000},"page":"2422","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["A Novel Highly Sensitive Method for Measuring Inflammatory Neural-Derived APC Activity in Glial Cell Lines, Mouse Brain and Human CSF"],"prefix":"10.3390","volume":"21","author":[{"given":"Valery","family":"Golderman","sequence":"first","affiliation":[{"name":"Department of Neurology, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv 6997801, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6104-606X","authenticated-orcid":false,"given":"Shany G.","family":"Gofrit","sequence":"additional","affiliation":[{"name":"Department of Neurology, The Chaim Sheba Medical Center, Ramat Gan 5266202, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nicola","family":"Maggio","sequence":"additional","affiliation":[{"name":"Department of Neurology, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv 6997801, Israel"},{"name":"Department of Neurology, The Chaim Sheba Medical Center, Ramat Gan 5266202, Israel"},{"name":"Talpiot Medical Leadership Program, The Chaim Sheba Medical Center, Ramat Gan 5266202, Israel"},{"name":"Sagol School of Neuroscience, Tel Aviv University, Tel Aviv 6997801, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Orna","family":"Gera","sequence":"additional","affiliation":[{"name":"Department of Neurology, The Chaim Sheba Medical Center, Ramat Gan 5266202, Israel"},{"name":"Department of Physical Therapy, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv 6997801, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexandra","family":"Gerasimov","sequence":"additional","affiliation":[{"name":"Department of Neurology, The Chaim Sheba Medical Center, Ramat Gan 5266202, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dar","family":"Laks","sequence":"additional","affiliation":[{"name":"Department of Neurology, The Chaim Sheba Medical Center, Ramat Gan 5266202, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joab","family":"Chapman","sequence":"additional","affiliation":[{"name":"Department of Neurology, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv 6997801, Israel"},{"name":"Department of Neurology, The Chaim Sheba Medical Center, Ramat Gan 5266202, Israel"},{"name":"Sagol School of Neuroscience, Tel Aviv University, Tel Aviv 6997801, Israel"},{"name":"Department of Physiology and Pharmacology, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv 6997801, Israel"},{"name":"Robert and Martha Harden Chair in Mental and Neurological Diseases, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv 6997801, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3373-8397","authenticated-orcid":false,"given":"Efrat","family":"Shavit-Stein","sequence":"additional","affiliation":[{"name":"Department of Neurology, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv 6997801, Israel"},{"name":"Department of Neurology, The Chaim Sheba Medical Center, Ramat Gan 5266202, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,3,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1016\/S1474-4422(15)70016-5","article-title":"Neuroinflammation in Alzheimer\u2019s disease","volume":"14","author":"Heneka","year":"2015","journal-title":"Lancet Neurol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"326","DOI":"10.1016\/j.bbrc.2014.09.068","article-title":"Inflammatory cytokines, interleukin-1 beta and tumor necrosis factor-alpha, upregulated in glioblastoma multiforme, raise the levels of CRYAB in exosomes secreted by U373 glioma cells","volume":"453","author":"Kore","year":"2014","journal-title":"Biochem. 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