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1.
Acta Neuropathol Commun ; 9(1): 144, 2021 08 26.
Article in English | MEDLINE | ID: mdl-34446086

ABSTRACT

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease involving progressive degeneration of upper and lower motor neurons. The pattern of lower motor neuron loss along the spinal cord follows the pattern of deposition of phosphorylated TDP-43 aggregates. The blood-spinal cord barrier (BSCB) restricts entry into the spinal cord parenchyma of blood components that can promote motor neuron degeneration, but in ALS there is evidence for barrier breakdown. Here we sought to quantify BSCB breakdown along the spinal cord axis, to determine whether BSCB breakdown displays the same patterning as motor neuron loss and TDP-43 proteinopathy. Cerebrospinal fluid hemoglobin was measured in living ALS patients (n = 87 control, n = 236 ALS) as a potential biomarker of BSCB and blood-brain barrier leakage. Cervical, thoracic, and lumbar post-mortem spinal cord tissue (n = 5 control, n = 13 ALS) were then immunolabelled and semi-automated imaging and analysis performed to quantify hemoglobin leakage, lower motor neuron loss, and phosphorylated TDP-43 inclusion load. Hemoglobin leakage was observed along the whole ALS spinal cord axis and was most severe in the dorsal gray and white matter in the thoracic spinal cord. In contrast, motor neuron loss and TDP-43 proteinopathy were seen at all three levels of the ALS spinal cord, with most abundant TDP-43 deposition in the anterior gray matter of the cervical and lumbar cord. Our data show that leakage of the BSCB occurs during life, but at end-stage disease the regions with most severe BSCB damage are not those where TDP-43 accumulation is most abundant. This suggests BSCB leakage and TDP-43 pathology are independent pathologies in ALS.


Subject(s)
Amyotrophic Lateral Sclerosis/cerebrospinal fluid , Amyotrophic Lateral Sclerosis/pathology , Blood-Brain Barrier/pathology , Cerebrospinal Fluid Leak/pathology , Motor Neurons/pathology , Spinal Cord/pathology , Adult , Aged , Aged, 80 and over , Blood-Brain Barrier/metabolism , Cerebrospinal Fluid Leak/metabolism , Female , Hemoglobins/cerebrospinal fluid , Humans , Male , Middle Aged , Motor Neurons/metabolism , Spinal Cord/metabolism
2.
J Cereb Blood Flow Metab ; 41(11): 3000-3015, 2021 11.
Article in English | MEDLINE | ID: mdl-34102922

ABSTRACT

Secondary brain injury after aneurysmal subarachnoid hemorrhage (SAH-SBI) contributes to poor outcomes in patients after rupture of an intracranial aneurysm. The lack of diagnostic biomarkers and novel drug targets represent an unmet need. The aim of this study was to investigate the clinical and pathophysiological association between cerebrospinal fluid hemoglobin (CSF-Hb) and SAH-SBI. In a cohort of 47 patients, we collected daily CSF-samples within 14 days after aneurysm rupture. There was very strong evidence for a positive association between spectrophotometrically determined CSF-Hb and SAH-SBI. The accuracy of CSF-Hb to monitor for SAH-SBI markedly exceeded that of established methods (AUC: 0.89 [0.85-0.92]). Temporal proteome analysis revealed erythrolysis accompanied by an adaptive macrophage response as the two dominant biological processes in the CSF-space after aneurysm rupture. Ex-vivo experiments on the vasoconstrictive and oxidative potential of Hb revealed critical inflection points overlapping CSF-Hb thresholds in patients with SAH-SBI. Selective depletion and in-solution neutralization by haptoglobin or hemopexin efficiently attenuated the vasoconstrictive and lipid peroxidation activities of CSF-Hb. Collectively, the clinical association between high CSF-Hb levels and SAH-SBI, the underlying pathophysiological rationale, and the favorable effects of haptoglobin and hemopexin in ex-vivo experiments position CSF-Hb as a highly attractive biomarker and potential drug target.


Subject(s)
Biomarkers/cerebrospinal fluid , Brain Ischemia/etiology , Hemoglobins/cerebrospinal fluid , Subarachnoid Hemorrhage/complications , Vasospasm, Intracranial/etiology , Adult , Aged , Cerebrospinal Fluid/chemistry , Female , Humans , Male , Middle Aged
3.
Front Immunol ; 11: 228, 2020.
Article in English | MEDLINE | ID: mdl-32210955

ABSTRACT

Intraventricular hemorrhage (IVH) is a frequent complication of prematurity that is associated with high neonatal mortality and morbidity. IVH is accompanied by red blood cell (RBC) lysis, hemoglobin (Hb) oxidation, and sterile inflammation. Here we investigated whether extracellular Hb, metHb, ferrylHb, and heme contribute to the inflammatory response after IVH. We collected cerebrospinal fluid (CSF) (n = 20) from premature infants with grade III IVH at different time points after the onset of IVH. Levels of Hb, metHb, total heme, and free heme were the highest in CSF samples obtained between days 0 and 20 after the onset of IVH and were mostly non-detectable in CSF collected between days 41 and 60 of post-IVH. Besides Hb monomers, we detected cross-linked Hb dimers and tetramers in post-IVH CSF samples obtained in days 0-20 and 21-40, but only Hb tetramers were present in CSF samples obtained after 41-60 days. Vascular cell adhesion molecule-1 (VCAM-1) and interleukin-8 (IL-8) levels were higher in CSF samples obtained between days 0 and 20 than in CSF collected between days 41 and 60 of post-IVH. Concentrations of VCAM-1, intercellular adhesion molecule-1 (ICAM-1), and IL-8 strongly correlated with total heme levels in CSF. Applying the identified heme sources on human brain microvascular endothelial cells revealed that Hb oxidation products and free heme contribute to the inflammatory response. We concluded that RBC lysis, Hb oxidation, and heme release are important components of the inflammatory response in IVH. Pharmacological interventions targeting cell-free Hb, Hb oxidation products, and free heme could have potential to limit the neuroinflammatory response following IVH.


Subject(s)
Brain/pathology , Cerebral Intraventricular Hemorrhage/metabolism , Endothelial Cells/metabolism , Erythrocytes/pathology , Heme/cerebrospinal fluid , Hemoglobins/cerebrospinal fluid , Inflammation/metabolism , Premature Birth/metabolism , Female , Humans , Infant, Newborn , Infant, Premature , Intercellular Adhesion Molecule-1/cerebrospinal fluid , Interleukin-8/cerebrospinal fluid , Male , Neurogenic Inflammation , Oxidation-Reduction , Premature Birth/immunology , Vascular Cell Adhesion Molecule-1/cerebrospinal fluid
4.
Nutr Neurosci ; 21(1): 40-48, 2018 Jan.
Article in English | MEDLINE | ID: mdl-27499134

ABSTRACT

OBJECTIVES: Iron deficiency (ID) anemia leads to long-term neurodevelopmental deficits by altering iron-dependent brain metabolism. The objective of the study was to determine if ID induces metabolomic abnormalities in the cerebrospinal fluid (CSF) in the pre-anemic stage and to ascertain the aspects of abnormal brain metabolism affected. METHODS: Standard hematological parameters [hemoglobin (Hgb), mean corpuscular volume (MCV), transferrin (Tf) saturation, and zinc protoporphyrin/heme (ZnPP/H)] were compared at 2, 4, 6, 8, and 12 months in iron-sufficient (IS; n = 7) and iron-deficient (ID; n = 7) infant rhesus monkeys. Five CSF metabolite ratios were determined at 4, 8, and 12 months using 1H NMR spectroscopy at 16.4 T and compared between groups and in relation to hematologic parameters. RESULTS: ID infants developed ID (Tf saturation < 25%) by 4 months of age and all became anemic (Hgb < 110 g/L and MCV < 60 fL) at 6 months. Their heme indices normalized by 12 months. Pyruvate/glutamine and phosphocreatine/creatine (PCr/Cr) ratios in CSF were lower in the ID infants by 4 months (P < 0.05). The PCr/Cr ratio remained lower at 8 months (P = 0.02). ZnPP/H, an established blood marker of pre-anemic ID, was positively correlated with the CSF citrate/glutamine ratio (marginal correlation, 0.34; P < 0.001; family wise error rate = 0.001). DISCUSSION: Metabolomic analysis of the CSF is sensitive for detecting the effects of pre-anemic ID on brain energy metabolism. Persistence of a lower PCr/Cr ratio at 8 months, even as hematological measures demonstrated recovery from anemia, indicate that the restoration of brain energy metabolism is delayed. Metabolomic platforms offer a useful tool for early detection of the impact of ID on brain metabolism in infants.


Subject(s)
Anemia, Iron-Deficiency/cerebrospinal fluid , Brain/metabolism , Iron/cerebrospinal fluid , Metabolomics , Animals , Animals, Newborn , Diet , Dietary Carbohydrates/administration & dosage , Dietary Fats/administration & dosage , Dietary Fiber/administration & dosage , Dietary Proteins/administration & dosage , Female , Hemoglobins/cerebrospinal fluid , Macaca mulatta , Magnetic Resonance Spectroscopy , Micronutrients/administration & dosage , Micronutrients/cerebrospinal fluid , Protoporphyrins/cerebrospinal fluid , Specimen Handling , Transferrin/cerebrospinal fluid
5.
Acta Neurochir Suppl ; 120: 51-4, 2015.
Article in English | MEDLINE | ID: mdl-25366599

ABSTRACT

Rapid and effective clearance of cell-free haemoglobin after subarachnoid haemorrhage (SAH) is important to prevent vasospasm and neurotoxicity and improve long-term outcome. Haemoglobin is avidly bound by haptoglobin, and the complex is cleared by CD163 expressed on the membrane surface of macrophages. We studied the kinetics of haemoglobin and haptoglobin in cerebrospinal fluid after SAH. We show that haemoglobin levels rise gradually after SAH. Haptoglobin levels rise acutely with aneurysmal rupture as a result of injection of blood into the subarachnoid space. Although levels decline as haemoglobin scavenging occurs, complete depletion of haptoglobin does not occur and levels start rising again, indicating saturation of CD163 sites available for haptoglobin-haemoglobin clearance. In a preliminary neuropathological study we demonstrate that meningeal CD163 expression is upregulated after SAH, in keeping with a proinflammatory state. However, loss of CD163 occurs in meningeal areas with overlying blood compared with areas without overlying blood. Becauses ADAM17 is the enzyme responsible for shedding membrane-bound CD163, its inhibition may be a potential therapeutic strategy after SAH.


Subject(s)
Antigens, CD/cerebrospinal fluid , Antigens, Differentiation, Myelomonocytic/cerebrospinal fluid , Haptoglobins/cerebrospinal fluid , Hemoglobins/cerebrospinal fluid , Subarachnoid Hemorrhage/cerebrospinal fluid , Vasospasm, Intracranial/cerebrospinal fluid , Humans , Kinetics , Protein Binding , Receptors, Cell Surface , Subarachnoid Hemorrhage/complications , Tissue Banks , Up-Regulation , Vasospasm, Intracranial/etiology , Vasospasm, Intracranial/prevention & control
6.
Biomark Med ; 8(3): 387-94, 2014.
Article in English | MEDLINE | ID: mdl-24712430

ABSTRACT

AIM: The purpose of this study was to establish the influence of centrifugation and protease activity on the cerebrospinal fluid (CSF) concentrations of DJ-1 and hemoglobin. MATERIALS & METHODS: The concentrations of DJ-1 and hemoglobin were determined in 12 (DJ-1) and six (hemoglobin) pairs of CSF samples, with one sample being stored without centrifugation and the other being centrifuged at 2000 × g before storage. The DJ-1 concentration was also determined in centrifuged and uncentrifuged CSF containing protease inhibitors and compared with values determined in centrifuged and uncentrifuged CSF samples without protease inhibitors. Furthermore, specific protein concentrations were determined in CSF from two groups, each comprising 23 patients with Parkinson's disease. In one group the CSF was centrifuged at 1300-1800 × g, 4°C, 10 min, and in the other at 2000 × g, 4°C, 10 min. RESULTS: Centrifugation at 2000 × g resulted in significantly lower CSF DJ-1 concentrations compared with no centrifugation and centrifugation at a lower g-force. There was a significant difference in the hemoglobin concentration between centrifuged and uncentrifuged CSF. In all centrifuged samples the hemoglobin concentration was <200 ng/ml including blood contaminated samples centrifuged at 2000 × g. When a protease inhibitor cocktail was added to the CSF prior to centrifugation, the DJ-1 concentration was significantly higher. CONCLUSION: Preanalytical factors such as centrifugation and protease inhibition must be carefully controlled when handling CSF for analysis of DJ-1 and other biomarkers, as DJ-1 was influenced by blood contamination, centrifugation and protease activity.


Subject(s)
Enzyme-Linked Immunosorbent Assay , Intracellular Signaling Peptides and Proteins/cerebrospinal fluid , Oncogene Proteins/cerebrospinal fluid , Aged , Biomarkers/cerebrospinal fluid , Centrifugation , Erythrocyte Count , Female , Hemoglobins/cerebrospinal fluid , Humans , Male , Middle Aged , Parkinson Disease/metabolism , Parkinson Disease/pathology , Protease Inhibitors/chemistry , Protein Deglycase DJ-1
7.
J R Coll Physicians Edinb ; 43(3): 200-6, 2013.
Article in English | MEDLINE | ID: mdl-24087797

ABSTRACT

BACKGROUND: Patients with suspected subarachnoid haemorrhage, a normal noncontrast computed tomography (CT) and cerebrospinal fluid (CSF) evidence of haemoglobin breakdown products often undergo CT angiography (CTA). If this is normal, then invasive catheter angiography may be offered. In current clinical practice, haemoglobin breakdown products are detected by spectrophotometry rather than visible xanthochromia, and CTA is performed on multidetector scanners. The aim of this study was to determine if such patients should still have a catheter angiography, given the associated risks. METHODS: Patients positive for CSF spectrophotometry (n=26) were retrospectively identified from the clinical biochemistry information system and imaging data from the electronic radiology records were reviewed. Discharge letters were consulted to relate the biochemistry and radiology results to the final diagnosis. RESULTS: 15 patients with CT angiography were found. Nine patients had normal CT angiography. No causative aneurysms had been missed. One patient had small, coincidental aneurysms missed on initial reading of the CTA. CONCLUSION: The likelihood of a clinically significant aneurysm in a patient who is CT negative, lumbar puncture positive and CTA negative is low. Double reporting of negative CT angiograms may be advisable.


Subject(s)
Cerebral Angiography , Hemoglobins/cerebrospinal fluid , Subarachnoid Hemorrhage/diagnostic imaging , Aged , Cerebral Angiography/methods , Humans , Retrospective Studies , Spectrophotometry , Subarachnoid Hemorrhage/cerebrospinal fluid , Tomography, X-Ray Computed/methods
8.
Clin Neurol Neurosurg ; 115(7): 965-70, 2013 Jul.
Article in English | MEDLINE | ID: mdl-23107165

ABSTRACT

OBJECTIVE: Subarachnoid clots play an important role in development of delayed vasospasm after subarachnoid hemorrhage (SAH). The purpose of this study was to compare clearance of subarachnoid clots using external ventricular drainage (EVD) or lumbar drainage (LD) after Guglielmi detachable coil (GDC) embolization for aneurysmal SAH. METHODS: The subjects were 51 treated with GDC coil embolization for aneurysmal Fisher group 3 SAH within 72 h of ictus. Software-based volumetric quantification of the subarachnoid clots was performed on CT scans and the hemoglobin (Hb) level was measured in CSF drained from each catheter. RESULTS: Clearance of subarachnoid clots was more rapid in patients treated with LD (n=34) compared to those treated with EVD (n=17). The Hb level in CSF was significantly higher in the LD group on Days 4-5 after onset of SAH (P<0.05), but was higher in the EVD group on Days 8-9. The incidence of symptomatic vasospasm did not differ between the two groups. The rate of occurrence of a new low density area on CT scans was higher in patients treated with EVD, but not significantly higher than the rate in the LD group. CONCLUSION: GDC embolization followed by lumbar drainage accelerates the reduction of subarachnoid clots, but EVD may contribute to stasis of hemorrhage within subarachnoid spaces.


Subject(s)
Drainage/methods , Embolization, Therapeutic/instrumentation , Embolization, Therapeutic/methods , Subarachnoid Hemorrhage/surgery , Subarachnoid Space/pathology , Adult , Aged , Aged, 80 and over , Cerebral Ventricles , Cohort Studies , Data Interpretation, Statistical , Female , Hemoglobins/cerebrospinal fluid , Humans , Lumbosacral Region , Male , Middle Aged , Neurosurgical Procedures , Paralysis/etiology , Subarachnoid Hemorrhage/cerebrospinal fluid , Tomography, X-Ray Computed , Treatment Outcome , Ultrasonography, Doppler, Transcranial , Vasospasm, Intracranial/etiology
9.
Arch Neurol ; 69(11): 1445-52, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22925882

ABSTRACT

OBJECTIVE: To assess the ability of 5 cerebrospinal fluid(CSF) biomarkers to differentiate between common dementia and parkinsonian disorders. DESIGN: A cross-sectional, clinic-based study. PARTICIPANTS: Cerebrospinal fluid samples (N=453) were obtained from healthy individuals serving as controls and from patients with Parkinson disease (PD), PD with dementia(PDD), dementia with Lewy bodies (DLB), Alzheimer disease (AD), progressive supranuclear palsy(PSP), multiple system atrophy (MSA), or corticobasal degeneration (CBD). SETTING: Neurology and memory disorder clinics. MAIN OUTCOME MEASURES: Cerebrospinal fluid biomarker levels in relation to clinical diagnosis. RESULTS: Cerebrospinal fluid levels of -synuclein were decreased in patients with PD, PDD, DLB, and MSA but increased in patients with AD. Cerebrospinal fluid levels of α-amyloid 1-42 were decreased in DLB and even further decreased in AD. Cerebrospinal fluid levels of total tau and hyperphosphorylated tau were increased in AD. Multivariate analysis revealed that these biomarkers could differentiate AD from DLB and PDD with an area under the curve of 0.90, with -synuclein and total tau contributing most to the model. Cerebrospinal fluid levels of neurofilament light chain were substantially increased in atypical parkinsonian disorders (ie, PSP, MSA,and CBD), and multivariate analysis revealed that the level of neurofilament light chain alone could differentiate PD from atypical parkinsonian disorders, with an area under the curve of 0.93. CONCLUSIONS: Ascertainment of the -synuclein level in CSF somewhat improves the differential diagnosis of AD vs DLB and PDD when combined with established AD biomarkers.The level of neurofilament light chain alone may differentiate PD from atypical parkinsonian disorders.


Subject(s)
Biomarkers/cerebrospinal fluid , Dementia/cerebrospinal fluid , Dementia/diagnosis , Parkinson Disease/cerebrospinal fluid , Parkinson Disease/diagnosis , Aged , Aged, 80 and over , Amyloid beta-Peptides/cerebrospinal fluid , Cross-Sectional Studies , Dementia/complications , Diagnosis, Differential , Enzyme-Linked Immunosorbent Assay , Female , Hemoglobins/cerebrospinal fluid , Humans , Male , Middle Aged , Multiple System Atrophy/cerebrospinal fluid , Multiple System Atrophy/diagnosis , Neurodegenerative Diseases/cerebrospinal fluid , Neurodegenerative Diseases/diagnosis , Neuropsychological Tests , Parkinson Disease/complications , Psychiatric Status Rating Scales , Severity of Illness Index , Statistics, Nonparametric , Supranuclear Palsy, Progressive/cerebrospinal fluid , Supranuclear Palsy, Progressive/diagnosis , alpha-Synuclein/cerebrospinal fluid , tau Proteins/cerebrospinal fluid
10.
J Chromatogr A ; 1267: 170-7, 2012 Dec 07.
Article in English | MEDLINE | ID: mdl-22841664

ABSTRACT

A CE-tandem MS method was optimised and validated for selective and specific determination of LVV- and VV-hemorphin-7 peptides in cerebrospinal fluid. These two small peptides originate from haemoglobin beta chains. They possess relevant biological activity and recently a potential biomarker role in posterior cranial fossa paediatric brain tumour disease was evidenced. The separation was optimised using formic acid as background electrolyte and a water/methanol mixture, containing 0.1% (v/v) formic acid, as sheath liquid. The two peptides, differing in only one amino acid of the sequence at the N-terminal side were baseline separated in less than 15 min. The method allowed a very reduced and rapid sample pretreatment and was successfully applied to hemorphins determination in patient samples without matrix interferences. The method successfully passed bioanalytical validation showing linearity, accuracy and precision data on cerebrospinal fluid matrix within the acceptable values. The analysis of cerebrospinal fluid of patients affected by different posterior cranial fossa tumour forms confirmed our previous findings showing the absence of hemorphins in the pre-surgical cerebrospinal fluid and their presence in the post-ones and controls. The present method saves costs and time due to capillary electrophoresis miniaturisation and to the absence of chromatographic column and gradient elution and allows numerous injections per sample starting from few microlitres of cerebrospinal fluid.


Subject(s)
Brain Neoplasms/cerebrospinal fluid , Electrophoresis, Capillary/methods , Hemoglobins/cerebrospinal fluid , Peptide Fragments/cerebrospinal fluid , Tandem Mass Spectrometry/methods , Brain Neoplasms/diagnosis , Child , Child, Preschool , Female , Humans , Male
11.
J Clin Neurosci ; 19(7): 961-4, 2012 Jul.
Article in English | MEDLINE | ID: mdl-22595353

ABSTRACT

Intraventricular hemorrhage (IVH) occurring after spontaneous intracerebral hemorrhage (ICH) is an independent risk factor for mortality. The use of intraventricular urokinase (Uk) to reduce intraventricular blood clot volume and improve outcome was investigated. Patients with IVH requiring external ventricular drainage were recruited and randomized into a double-blind placebo controlled study. Assessments of collected cerebrospinal fluid (CSF) haemoglobin (Hb) and serial CT scans were performed. The study outcomes were: infection rates, length of stay in the intensive care unit, survival, National Institutes of Health Stroke Scale score; and modified Rankin Scale scores. Our results showed an increase in both the drained CSF Hb concentration in patients treated with Uk compared to placebo and in the rate of resolution clot volume. No differences were found in the other outcome measures but there was a trend towards lowered mortality in the group treated with Uk. Therefore, intraventricular Uk resulted in faster resolution of IVH with no adverse events.


Subject(s)
Cerebral Hemorrhage/drug therapy , Fibrinolytic Agents/therapeutic use , Urokinase-Type Plasminogen Activator/therapeutic use , Aged , Analysis of Variance , Cerebral Hemorrhage/cerebrospinal fluid , Cerebral Hemorrhage/diagnostic imaging , Cerebral Hemorrhage/mortality , Double-Blind Method , Female , Hemoglobins/cerebrospinal fluid , Humans , Injections, Intraventricular , Kaplan-Meier Estimate , Male , Middle Aged , Pilot Projects , Severity of Illness Index , Statistics, Nonparametric , Time Factors , Tomography, X-Ray Computed , Treatment Outcome
12.
Proteomics ; 12(13): 2158-66, 2012 Jul.
Article in English | MEDLINE | ID: mdl-22623401

ABSTRACT

Posterior cranial fossa is the most frequent location of pediatric brain tumors. Its diagnosis is currently performed by postsurgery histopathology and the identification of biomarkers in cerebrospinal fluid (CSF) could provide a less invasive tool. Patient CSF was collected during surgery before the tumor removal (PRE-CSF) and 6 days after the resection (POST-CSF) and analyzed by top down LC-MS proteomics for comparison. The PRE-CSFs generally exhibited a less complex LC-MS profile than the relative POST-CSFs suggesting a suppressive role of the tumor toward proteins and peptides production or release. Particularly, a panel of peptides, identified as alpha- and beta-hemoglobin chains fragments, were generally absent in the PRE-CSF and present in the POST ones independently from contaminant blood hemoglobin. Among them, the LVV- and VV-hemorphin-7 showed the most repeatable trend and with a few remarkable exceptions: their unusual absence in POST surgery CSF was in fact interestingly correlated to the presence of tumor in the patient despite surgery due to metastases or to subtotal resection. These results ascribed a relevant biological role to LVV- and VV-h7 peptides in the disease and a strong potential as biomarkers. Their analysis in POST surgery CSF could be used to predict patient prognosis.


Subject(s)
Biomarkers, Tumor/cerebrospinal fluid , Brain Neoplasms/cerebrospinal fluid , Hemoglobins/cerebrospinal fluid , Peptide Fragments/cerebrospinal fluid , Proteomics/methods , Adolescent , Amino Acid Sequence , Biomarkers, Tumor/chemistry , Brain Neoplasms/pathology , Child , Child, Preschool , Cranial Fossa, Posterior/pathology , Female , Hemoglobins/chemistry , Humans , Infant , Infant, Newborn , Male , Molecular Sequence Data , Peptide Fragments/chemistry
13.
J Neurochem ; 121(5): 785-92, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22380637

ABSTRACT

Delayed cerebral ischemia resulting from extracellular hemoglobin is an important determinant of outcome in subarachnoid hemorrhage. Hemoglobin is scavenged by the CD163-haptoglobin system in the circulation, but little is known about this scavenging pathway in the human CNS. The components of this system were analyzed in normal cerebrospinal fluid and after subarachnoid hemorrhage. The intrathecal presence of the CD163-haptoglobin-hemoglobin scavenging system was unequivocally demonstrated. The resting capacity of the CD163-haptoglobin-hemoglobin system in the normal CNS was 50 000-fold lower than that of the circulation. After subarachnoid hemorrhage, the intrathecal CD163-haptoglobin-hemoglobin system was saturated, as shown by the presence of extracellular hemoglobin despite detectable haptoglobin. Hemoglobin efflux from the CNS was evident, enabling rescue hemoglobin scavenging by the systemic circulation. Therefore, the CNS is not capable of dealing with significant intrathecal hemolysis. Potential therapeutic options to prevent delayed cerebral ischemia ought to concentrate on augmenting the capacity of the intrathecal CD163-haptoglobin-hemoglobin scavenging system and strategies to encourage hemoglobin efflux from the brain.


Subject(s)
Antigens, CD/cerebrospinal fluid , Antigens, Differentiation, Myelomonocytic/cerebrospinal fluid , Haptoglobins/cerebrospinal fluid , Hemoglobins/cerebrospinal fluid , Subarachnoid Hemorrhage/cerebrospinal fluid , Brain Ischemia/cerebrospinal fluid , Brain Ischemia/epidemiology , Brain Ischemia/etiology , Enzyme-Linked Immunosorbent Assay , Female , Humans , Macrophages/immunology , Macrophages/metabolism , Male , Middle Aged , Receptors, Cell Surface , Subarachnoid Hemorrhage/complications , Subarachnoid Hemorrhage/immunology
14.
Neurosurgery ; 70(3): 602-9, 2012 Mar.
Article in English | MEDLINE | ID: mdl-21866060

ABSTRACT

BACKGROUND: Free radicals and lipid peroxidation are thought to be related to the vasospasm generation after subarachnoid hemorrhage (SAH). Plasma platelet-activating factor-acetyl hydrolase (PAF-AH) degrades phospholipids with an oxidatively modified fatty acyl chain. OBJECTIVE: To compare plasma PAF-AH activity and free forms of biomarker of lipid peroxidation in cerebrospinal fluid (CSF) between patients with and without symptomatic vasospasm (SVS) after SAH. METHODS: The identification of PAF-AH in CSF was performed by Western blotting. The genotype at position 279 of the plasma PAF-AH gene was determined. The activities of PAF-AH and the levels of free 8-iso-prostaglandin F2α (free isoPs), free hydroxyoctadecadienoic acid (free HODE), and free hydroxyeicosatetraenoic acid (free HETE) in CSF were measured. RESULTS: The PAF-AH in CSF was confirmed to be only the plasma type. The genotype of the plasma PAF-AH was not different between patients with and without SVS. Free isoPs, free HODE, and free HETE showed higher values in patients without SVS in 0 to 4 days and 5 to 9 days after SAH. The PAF-AH activity also was higher in patients without SVS in 0 to 4 days and 5 to 9 days after SAH. The associations between PAF-AH activity and free isoPs, and between PAF-AH activity and free HODE were significant. CONCLUSION: Oxidized lipids of lipoproteins and blood cell membranes produced by reactive oxygen species in CSF when SAH occurs may be the main source of lipid peroxidation. Plasma PAF-AH can hydrolyze oxidized phospholipids, and may attenuate the spreading of lipid peroxidation and participate in defense mechanisms against vasospasm after SAH.


Subject(s)
1-Alkyl-2-acetylglycerophosphocholine Esterase/blood , 1-Alkyl-2-acetylglycerophosphocholine Esterase/genetics , Lipid Peroxidation/physiology , Subarachnoid Hemorrhage , Vasospasm, Intracranial , Aged , Biomarkers/blood , Biomarkers/cerebrospinal fluid , Dinoprost/analogs & derivatives , Dinoprost/cerebrospinal fluid , Enzyme Activation/physiology , Fatty Acids, Unsaturated/cerebrospinal fluid , Female , Genotype , Hemoglobins/cerebrospinal fluid , Humans , Hydroxyeicosatetraenoic Acids/cerebrospinal fluid , Male , Middle Aged , Oxidative Stress/physiology , Phospholipids/metabolism , Subarachnoid Hemorrhage/blood , Subarachnoid Hemorrhage/cerebrospinal fluid , Subarachnoid Hemorrhage/genetics , Vasospasm, Intracranial/blood , Vasospasm, Intracranial/cerebrospinal fluid , Vasospasm, Intracranial/diagnosis
15.
IEEE Trans Biomed Eng ; 58(3): 773-6, 2011 Mar.
Article in English | MEDLINE | ID: mdl-21292590

ABSTRACT

In North America, an estimated 30,000 patients annually experience an aneurysmal subarachnoid hemorrhage (SAH). In approximately five percent of these patients, the hemorrhage is not visible on computerized tomography scans due to the inability to image blood at time intervals greater than 12 h post symptom onset. For these patients (many of which have experienced a sentinel hemorrhage that is a precursor to a more significant rupture), a method is needed for accurately analyzing cerebral spinal fluid (CSF) for evidence of SAH. Further, it is necessary to differentiate blood associated with the SAH from blood associated with the spinal tap procedure. This letter presents a point-of-care device that is capable of performing such an analysis. The stand-alone prototype device uses commercially available embedded system components to implement a point-of-care device that is capable of collecting and analyzing optical absorbance spectra. A mathematical model for the hemorrhagic CSF sample is then developed by using a partial-least-squares-regression-based regression methodology that is able to differentiate between SAH and blood associated with the spinal tap. This differentiation is achieved by quantifying bilirubin (associated with the breakdown of old blood) in the CSF. Initial testing on the prototype device suggests that the device is able to quantify bilirubin in the presence of hemoglobin over concentrations ranges that are clinically relevant to the patient population of interest.


Subject(s)
Bilirubin/cerebrospinal fluid , Point-of-Care Systems , Spectrum Analysis/instrumentation , Subarachnoid Hemorrhage/cerebrospinal fluid , Algorithms , Hemoglobins/cerebrospinal fluid , Humans , Least-Squares Analysis , Models, Biological , Spinal Puncture
16.
Article in English | MEDLINE | ID: mdl-22254236

ABSTRACT

Subarachnoid hemorrhage (SAH) is a dangerous neurological event with a very short time window for early diagnosis. Clinical diagnoses performed in a lab seek to quantify bilirubin in cerebrospinal fluid (CSF) as a biomarker for SAHs; however laboratory assays suffer from lengthy protocols, interference from hemoglobin, and the availability of expertise. Substantial improvements in the determination of bilirubin concentration in the presence of hemoglobin in CSF are demonstrated in this work. Concentration estimates within 15% for bilirubin in the range of 0.2 to 1.6 mg /dl were determined for CSF samples containing fresh hemoglobin concentrations ranging from 0.05 to 0.25 g/dl. To demonstrate extensibility of the system with respect to more complete mock SAH samples, sample sets with one additional species of both hemoglobin and bilirubin, methemoglobin and alpha-bilirubin, respectively, were tested and yielded results within 25% of actual values, as measured by standard chemical assays of preparations prior to mixing.


Subject(s)
Algorithms , Artifacts , Bilirubin/cerebrospinal fluid , Data Interpretation, Statistical , Hemoglobins/cerebrospinal fluid , Spectrophotometry, Ultraviolet/methods , Complex Mixtures/cerebrospinal fluid , Humans , Least-Squares Analysis , Reproducibility of Results , Sensitivity and Specificity , Spectrophotometry, Ultraviolet/instrumentation
17.
Clin Chem Lab Med ; 45(1): 112-3, 2007.
Article in English | MEDLINE | ID: mdl-17243927

ABSTRACT

BACKGROUND: Among other methods, trichloroacetic acid precipitation is used to quantify total protein in cerebrospinal fluid (CSF). METHODS: We analyzed the influence of hemoglobin on total protein concentration assayed by the trichloroacetic acid method and compared the results to the benzethonium chloride method. RESULTS: Four CSF samples were spiked with different amounts of hemoglobin, leading to overestimation of protein concentration when assayed by the trichloroacetic acid method. Using the benzethonium chloride method, measurement of protein concentration was minimally disturbed. In addition, albumin and total protein concentrations were measured in 135 clinical samples. The total protein/albumin ratio remained constant when protein was measured with the benzethonium chloride method, while ratios increased when protein was assayed by the trichloroacetic acid method. CONCLUSIONS: Strong interference by hemoglobin leads to overestimation of the total protein concentration in CSF when assayed by the trichloroacetic acid method and may lead to false conclusions when evaluating the blood-brain barrier.


Subject(s)
Cerebrospinal Fluid Proteins/analysis , Hemoglobins/cerebrospinal fluid , Trichloroacetic Acid , Benzethonium , Chemical Precipitation
19.
Am J Obstet Gynecol ; 193(3 Pt 2): 957-64, 2005 Sep.
Article in English | MEDLINE | ID: mdl-16157094

ABSTRACT

OBJECTIVE: Preeclampsia is an idiopathic multisystem disorder specific to human pregnancy. This study used proteomic analysis of cerebrospinal fluid (CSF) to identify protein biomarkers characteristic of preeclampsia and related to its severity. STUDY DESIGN: CSF was collected from women diagnosed clinically with severe preeclampsia (sPE: n = 7), mild preeclampsia (mPE: n = 8), and normotensive controls (CRL: n = 8). Samples were subjected to proteomic analysis using surface-enhanced laser desorption/ionization time-of-flight (SELDI-TOF) mass spectroscopy. A discriminative proteomic biomarker profile was extracted by applying Mass Restricted analysis, and a Preeclampsia Proteomic Biomarker (PPB) score developed based on the presence or absence of four discriminatory protein peaks in individual CSF SELDI tracings. In-gel tryptic digests, Western blot analysis, on-chip immunoassays, ELISA, and spectral analysis were used to identify the biomarkers composing the PPB score. RESULTS: PPB score distinguished patients with a clinical diagnosis of sPE from mPE and CRLs. (PPB median [range]: sPE: 4 [0-4] vs mPE: 1 [0-1] vs CRL: 0 [0-0]; P < 0.001). PPB scores were unaffected by parity, magnesium seizure prophylaxis, CSF leukocyte counts, and total protein content. Proteomic identification techniques matched the discriminatory protein peaks to the alpha- and beta-hemoglobin chains. ELISA confirmed that women diagnosed clinically with sPE had significantly higher CSF hemoglobin concentrations than women with mPE or CRL (median [range]: sPE: 6.6 [0.0-10.3] microg/mL vs mPE: 0 [0-1.3] microg/mL vs CRL: 0 [0-0] microg/mL; P < 0.001). CONCLUSION: Proteomic analysis of CSF can accurately distinguish sPE from both mPE and CRL. Patients with sPE have nanomolar amounts of free hemoglobin in their CSF. Further studies are needed to confirm these observations and determine their physiologic implications.


Subject(s)
Hemoglobins/cerebrospinal fluid , Pre-Eclampsia/cerebrospinal fluid , Biomarkers/analysis , Female , Humans , Mass Spectrometry , Pregnancy , Protein Array Analysis , Proteomics
20.
Klin Lab Diagn ; (8): 5-7, 2002 Aug.
Article in Russian | MEDLINE | ID: mdl-12362644

ABSTRACT

Erythrocytes, extracellular hemoglobin and bilirubin level, a quantity of medium mass molecules and catalase activity were determined in the spinal fluid of 32 patient with closed craniocerebral injury of the various degree 24 hours after accident. The extracellular hemoglobin and the bilirubin level were shown to appear in cases with cerebral contusion in association with bleeding only, failed to reflect state severity degree at the early stage after injury. As the number of medium mass molecules increases significantly only in the patients with severe contusions, this index can be used for description of the severity of the process and for evaluation of the deterioration of the clearance of proteolytic products resulting in endogenous intoxication of the central nervous system. The spinal catalytic enzyme activity reflects the severity of the closed cerebrocranial trauma most objectively and can be used as a subarachnoid hemorrhage marker.


Subject(s)
Head Injuries, Closed/cerebrospinal fluid , Bilirubin/cerebrospinal fluid , Biomarkers/cerebrospinal fluid , Catalase/cerebrospinal fluid , Erythrocyte Count , Head Injuries, Closed/diagnosis , Hemoglobins/cerebrospinal fluid , Humans
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