Mass spectrometry‐based method for quantification of nimodipine and glutamate in cerebrospinal fluid. Pilot study with patients after aneurysmal subarachnoid haemorrhage

Introduction. High-mobility group box 1 (HMGB1) is an alarmin with proinflammatory potential determined by redox status of the cysteines at position 23 and 45. It may also play a role as a biomarker in biological fluids. The aim of this study was the identification of different HMGB1 redox forms in cerebrospinal fluid (CSF) obtained from subarachnoid hemorrhage patients. Material and Methods. 6 CSF samples were collected from aneurysmal subarachnoid haemorrhage patients. Commercially available HMGB1 isoforms served as a positive control. Immunoprecipitation and electrophoretic isolation of HMGB1 protein were performed, then both CSF and control were analyzed using mass spectrometry technique. To distinguish between fully reduced (thiol group at C23 and C45) and disulfide (disulfide bond connecting C23 and C45) HMGB1 forms, top-down sequencing of the spectra was performed. Results. Top-down sequencing analysis allowed to distinguish between HMGB1 isoforms only in commercially available standard without preceding immunoprecipitation and electrophoresis. MALDI spectra differ i.e. on the fully reduced HMGB1 spectrum fragmentation occurs before and beyond C22, which is not present on the disulfide HMGB1 spectrum. Analysis of HMGB1 isolated from CSF obtained from subarachnoid hemorrhage patients gave no results. Conclusions. Top-down sequencing enables to distinguish between redox forms of HMGB1. Electrophoresis and tryptic digestion cannot precede mass spectrometry analysis of redox forms of HMGB1 due to the reduction of disulfide bonds during these processes. Preferred method of isolation of HMGB1 for direct analysis using top-down sequencing mustn’t include protein digestion or degradation.

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Correction: Soluble Toll-Like Receptors 2 and 4 in Cerebrospinal Fluid of Patients with Acute Hydrocephalus following Aneurysmal Subarachnoid Haemorrhage

PLoS ONE ◽

10.1371/journal.pone.0157853 ◽

2016 ◽

Vol 11 (6) ◽

pp. e0157853

Author(s):

Bartosz Sokół ◽

Norbert Wąsik ◽

Roman Jankowski ◽

Marcin Hołysz ◽

Barbara Więckowska ◽

...

Keyword(s):

Cerebrospinal Fluid ◽

Subarachnoid Haemorrhage ◽

Aneurysmal Subarachnoid Haemorrhage ◽

Toll Like Receptors ◽

Acute Hydrocephalus

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Monitoring fibrinolytic activity in the cerebrospinal fluid after aneurysmal subarachnoid haemorrhage: a guide to the risk of rebleeding?

Journal of Neurology Neurosurgery & Psychiatry ◽

10.1136/jnnp.43.2.175 ◽

1980 ◽

Vol 43 (2) ◽

pp. 175-181 ◽

Cited By ~ 18

Author(s):

R S Maurice-Williams ◽

Y B Gordon ◽

A Sykes

Keyword(s):

Cerebrospinal Fluid ◽

Subarachnoid Haemorrhage ◽

Fibrinolytic Activity ◽

Aneurysmal Subarachnoid Haemorrhage

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Increase of Soluble RAGE in Cerebrospinal Fluid following Subarachnoid Haemorrhage

BioMed Research International ◽

10.1155/2017/7931534 ◽

2017 ◽

Vol 2017 ◽

pp. 1-7 ◽

Cited By ~ 3

Author(s):

Bartosz Sokół ◽

Norbert Wąsik ◽

Roman Jankowski ◽

Marcin Hołysz ◽

Witold Mańko ◽

...

Keyword(s):

Cerebrospinal Fluid ◽

Subarachnoid Haemorrhage ◽

Control Group ◽

Aneurysmal Subarachnoid Haemorrhage ◽

Pronounced Difference ◽

Significance Threshold ◽

Soluble Rage ◽

Inflammatory Mechanism ◽

Glycation End Products ◽

Csf Levels

Receptors for advanced glycation end-products (RAGE) mediate the inflammatory reaction that follows aneurysmal subarachnoid haemorrhage. Soluble RAGE (sRAGE) may function as a decoy receptor. The significance of this endogenous anti-inflammatory mechanism in subarachnoid haemorrhage (SAH) remains unknown. The present study aims to analyse sRAGE levels in the cerebrospinal fluid (CSF) of SAH patients. sRAGE levels were assayed by ELISA kit in 47 CSF samples collected on post-SAH days 0–3, 5–7, and 10–14 from 27 SAH patients with acute hydrocephalus. CSF levels of sRAGE were compared with a control group and correlated with other monitored parameters. In the control group, the CSF contained only a trace amount of sRAGE. By contrast, the CSF of 20 SAH patients collected on post-SAH days 0–3 was found to contain statistically significant higher levels of sRAGE (mean concentration 3.91 pg/mL, p<0.001). The most pronounced difference in CSF sRAGE levels between good and poor outcome patients was found on days 0–3 post-SAH but did not reach the significance threshold (p=0.234). CSF sRAGE levels did not change significantly during hospitalisation (p=0.868) and correlated poorly with treatment outcome, systemic inflammatory markers, and other monitored parameters. Our study revealed an early and constant increase of sRAGE level in the CSF of SAH patients.

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