0bb014d1111726c625a8d7e5d2a0513a pmc_10931830.pdf bcf459e7d09ecb734a454ca79ecd7cf5dfa7545d pmc_10931830.pdf 19775fd2ee023452eb5f5b85f2299ed893db2b96f7a968802b7bdcd3ff0eb697 pmc_10931830.pdf Title: Investigating the Role of Brain Natriuretic Peptide (BNP) and N-Terminal-proBNP in Thrombosis and Acute Ischemic Stroke Etiology Subject: The need for biomarkers for acute ischemic stroke (AIS) to understand the mechanisms implicated in pathological clot formation is critical. The levels of the brain natriuretic peptides known as brain natriuretic peptide (BNP) and NT-proBNP have been shown to be increased in patients suffering from heart failure and other heart conditions. We measured their expression in AIS clots of cardioembolic (CE) and large artery atherosclerosis (LAA) etiology, evaluating their location inside the clots, aiming to uncover their possible role in thrombosis. We analyzed 80 thrombi from 80 AIS patients in the RESTORE registry of AIS clots, 40 of which were of CE and 40 of LAA etiology. The localization of BNP and NT-BNP, quantified using immunohistochemistry and immunofluorescence, in AIS-associated white blood cell subtypes was also investigated. We found a statistically significant positive correlation between BNP and NT-proBNP expression levels (Spearman’s rho = 0.668 p < 0.0001 *). We did not observe any statistically significant difference between LAA and CE clots in BNP expression (0.66 [0.13–3.54]% vs. 0.53 [0.14–3.07]%, p = 0.923) or in NT-proBNP expression (0.29 [0.11–0.58]% vs. 0.18 [0.05–0.51]%, p = 0.119), although there was a trend of higher NT-proBNP expression in the LAA clots. It was noticeable that BNP was distributed throughout the thrombus and especially within platelet-rich regions. However, NT-proBNP colocalized with neutrophils, macrophages, and T-lymphocytes, suggesting its association with the thrombo-inflammatory process. Keywords: brain natriuretic peptide; BNP; NT-proBNP; stroke biomarkers; thrombus; acute ischemic stroke; stroke etiology Author: Rosanna Rossi, Duaa Jabrah, Andrew Douglas, James Prendergast, Abhay Pandit, Michael Gilvarry, Ray McCarthy, Petra Redfors, Annika Nordanstig, Turgut Tatlisumak, Erik Ceder, Dennis Dunker, Jeanette Carlqvist, István Szikora, Georgios Tsivgoulis, Klearchos Psychogios, John Thornton, Alexandros Rentzos, Katarina Jood, Jesus Juega and Karen M. Doyle Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Tue Mar 5 08:27:37 2024 CET ModDate: Tue Mar 5 08:40:12 2024 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 12 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 6869367 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- AIQCCL+VnURWPalladioL-Bold Type 1 Custom yes yes yes 10 0 FPGVTE+VnURWPalladioL Type 1 Custom yes yes yes 16 0 KRWGZA+URWPalladioL-Bold Type 1 Custom yes yes yes 21 0 ETQPPN+URWPalladioL-Roma Type 1 Custom yes yes yes 27 0 SEOVDY+URWPalladioL-Ital Type 1 Custom yes yes yes 32 0 IRWHZX+TimesNewRomanPSMT Type 1C WinAnsi yes yes no 48 0 CKUPDE+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 51 0 VIWIOF+CMSY10 Type 1 Builtin yes yes yes 71 0 HPHLBP+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 86 0 HPHLCA+PalatinoLinotype CID TrueType Identity-H yes yes yes 89 0 HPHLCB+PalatinoLinotype TrueType WinAnsi yes yes no 95 0 HPHLCD+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 98 0 HPHLBP+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 112 0 HPHLCB+PalatinoLinotype TrueType WinAnsi yes yes no 115 0 HPHLCD+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 118 0 HPHLLA+PalatinoLinotype TrueType MacRoman yes yes no 121 0 OVNEXW+EURM10 Type 1 Builtin yes yes yes 128 0 HPHLBP+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 140 0 HPHLCA+PalatinoLinotype CID TrueType Identity-H yes yes yes 143 0 HPHLCB+PalatinoLinotype TrueType WinAnsi yes yes no 149 0 HPHLCD+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 152 0 HPHLLA+PalatinoLinotype TrueType MacRoman yes yes no 155 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-05-07 04:19:59 CEST RepresentationInformation: pmc_10931830.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-03-20 02:30:44 CET Size: 6869367 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 335 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Results Destination: section.2 Children: Item: Title: Characteristics of the Patients Destination: subsection.2.1 Item: Title: BNP and NT-proBNP Expression Levels in AIS Clots Are Positively Correlated, and the Expression of Each Molecule Is Similar in LAA and CE Clots Destination: subsection.2.2 Item: Title: NT-ProBNP Expression in AIS Clots Is Associated with T-Lymphocytes, Neutrophils, and Macrophages Destination: subsection.2.3 Item: Title: Discussion Destination: section.3 Item: Title: Materials and Methods Destination: section.4 Children: Item: Title: Patient Cohort Destination: subsection.4.1 Item: Title: AIS Clot Collection and Processing Destination: subsection.4.2 Item: Title: Immunohistochemistry Staining Destination: subsection.4.3 Item: Title: Slide Scanning and Quantification Destination: subsection.4.4 Item: Title: Immunofluorescence Staining Destination: subsection.4.5 Item: Title: Statistical Analysis Destination: subsection.4.6 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: appendix.A. Info: Title: Investigating the Role of Brain Natriuretic Peptide (BNP) and N-Terminal-proBNP in Thrombosis and Acute Ischemic Stroke Etiology Author: Rosanna Rossi, Duaa Jabrah, Andrew Douglas, James Prendergast, Abhay Pandit, Michael Gilvarry, Ray McCarthy, Petra Redfors, Annika Nordanstig, Turgut Tatlisumak, Erik Ceder, Dennis Dunker, Jeanette Carlqvist, István Szikora, Georgios Tsivgoulis, Klearchos Psychogios, John Thornton, Alexandros Rentzos, Katarina Jood, Jesus Juega and Karen M. Doyle Subject: The need for biomarkers for acute ischemic stroke (AIS) to understand the mechanisms implicated in pathological clot formation is critical. The levels of the brain natriuretic peptides known as brain natriuretic peptide (BNP) and NT-proBNP have been shown to be increased in patients suffering from heart failure and other heart conditions. We measured their expression in AIS clots of cardioembolic (CE) and large artery atherosclerosis (LAA) etiology, evaluating their location inside the clots, aiming to uncover their possible role in thrombosis. We analyzed 80 thrombi from 80 AIS patients in the RESTORE registry of AIS clots, 40 of which were of CE and 40 of LAA etiology. The localization of BNP and NT-BNP, quantified using immunohistochemistry and immunofluorescence, in AIS-associated white blood cell subtypes was also investigated. We found a statistically significant positive correlation between BNP and NT-proBNP expression levels (Spearman’s rho = 0.668 p < 0.0001 *). We did not observe any statistically significant difference between LAA and CE clots in BNP expression (0.66 [0.13 3.54]% vs. 0.53 [0.14 3.07]%, p = 0.923) or in NT-proBNP expression (0.29 [0.11 0.58]% vs. 0.18 [0.05 0.51]%, p = 0.119), although there was a trend of higher NT-proBNP expression in the LAA clots. It was noticeable that BNP was distributed throughout the thrombus and especially within platelet-rich regions. However, NT-proBNP colocalized with neutrophils, macrophages, and T-lymphocytes, suggesting its association with the thrombo-inflammatory process. 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