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[Effect of teicoplanin and vancomycin on cerebrospinal fluid proteins of non-infected rabbits after suboccipital injection].

The IC inoculation of antibiotics into the CSF for therapeutic use could produce biological effects we should consider when analysing samples. To corroborate this assumption, we observed the effect of ICI T and V on the PL of the CSF of non infected rabbits. T and V were ICI as dosage of 1 mg/kg diluted in 0.2 ml of isotonic saline solution (ISS). ISS was also ICI alone. CSF samples were obtained before inoculation from 41 animals (T0) setting the normal PL. Other samples were obtained 2 (T2) and 4 hours (T4) after inoculation. PL were assayed in an Analyser Clinic Automatic (Du Pont). The statistical analysis was performed by the Kolomogorov-Smirnov Test, for comparison of samples from unknown and not necessarily similar distributions. Results were (mg/l) = PL at T0 = 0.20 +/- 0.08. At T2, levels were 0.6 +/- 0.41 (ISS), 0.73 +/- 0.27 (V) and 0.87 +/- 0.44 (T). At T4 they were 0.3 +/- 0.15 (ISS), 0.55 +/- 0.25 (V) and 0.78 +/- (T). Statistical differences (p less than 0.05) were demonstrated at T2 (T, V and ISS vs control at T0), at T4 = V, vs control at T0 but not between the two antibiotics nor between the two antibiotics and ISS, at any time. We conclude that IC inoculation of T and V and ISS increased significantly the CSF PL.

Animals↗

Cerebrospinal fluid proteins in patients with leucoaraiosis: possible abnormalities in blood-brain barrier function.

Some CSF protein abnormalities have been proposed as a possible marker for vascular dementia. We studied the CSF protein levels and albumin ratio in 21 patients (mean age 64.04 +/- 7.5) with progressive bilateral motor impairment, and a CT picture of leucoaraiosis. Seven of these patients also presented with dementia. Twenty-seven Alzheimer's disease patients (mean age 59.59 +/- 5.30) without leucoaraiosis were taken as controls. We also evaluated the correlations of the albumin ratio values with the diagnosis of dementia, the severity of cognitive impairment, the degree of cerebral atrophy and presence of infarcts on CT, and the abnormalities in CSF circulation, found on isotopic cisternography, in the leucoaraiosis group. After controlling for age and sex, the patients with leucoaraiosis showed greater CSF albumin levels (0.27 g/l +/- 0.11 vs. 0.21 g/l +/- 0.06; covariance analysis P = 0.066), CSF IgG values (4.68 mg/100 ml +/- 1.45 vs. 2.85 mg/100 ml +/- 1.03; covariance analysis P < 0.001), and a higher albumin ratio (0.0078 +/- 0.0027 vs. 0.0058 +/- 0.0019; covariance analysis P = 0.013) than those with Alzheimer's disease. The variations of these parameters were not apparently related to the presence of dementia in the leucoaraiosis group. A significantly higher albumin ratio was observed in patients with a slowed CSF circulation compared to those with normal CSF circulation (0.0086 +/- 0.0028 vs. 0.0059 +/- 0.0019; covariance analysis P = 0.05). We conclude that, independently from the presence of dementia, patients with leucoaraiosis have CSF abnormalities consistent with functional blood-brain barrier alterations.

Aged↗

Blood-brain barrier in chronic relapsing experimental allergic encephalomyelitis: a correlative study between cerebrospinal fluid protein concentrations and tracer leakage in the central nervous system.

Blood-brain barrier (BBB) permeability in chronic relapsing experimental allergic encephalomyelitis was studied morphologically in tracer studies with horseradish peroxidase (HRP) as well as by quantitative determination of HRP, albumin, and IgG in serum and cerebrospinal fluid (CSF). BBB damage was found to be localized in demyelinating plaques and in blood vessels with vasculitis. Actively demyelinating lesions showed massive increase in BBB permeability, whereas in inactive or remyelinated lesions BBB damage was either minimal or absent. Determination of serum proteins in the CSF of animals with severe disease and a high incidence of actively demyelinating lesions showed evidence of BBB damage (reduction of Q-albumin) and an IgG-index in the normal range. In animals with only inactive lesions the Q-albumin was normal, the IgG index, however, was elevated. This finding indicates intrathecal IgG synthesis. A correlation between morphologically visualized tracer leakage in the central nervous system (CNS) with serum protein concentrations in the CSF revealed that elevated CSF albumin is a reliable indicator for BBB damage in lesions, located near the inner or outer surface of the brain and spinal cord. However, singular focal lesions with BBB damage located in the depth of the CNS parenchyma may not be accompanied by CSF protein alterations. The invariable presence of BBB damage in active inflammatory demyelinating lesions and its absence in inactive plaques or in the unaffected nervous tissue may be important in therapy, not only in experimental allergic encephalomyelitis but also in multiple sclerosis (MS).

Animals↗

Apolipoprotein E and other cerebrospinal fluid proteins differentiate ante mortem variant Creutzfeldt-Jakob disease from ante mortem sporadic Creutzfeldt-Jakob disease.

The ability to perform an ante mortem differential diagnosis of Creutzfeld-Jakob disease (CJD) is aided by several clinical and molecular tests. There is a need for molecular tests which can reliably distinguish ante mortem variant CJD (vCJD) from ante mortem sporadic CJD (spCJD). A proteomics approach employing two-dimensional protein electrophoresis is applied to the study of ante mortem CSF samples obtained in collaboration with the CJD Surveillance Unit and the National Hospital for Neurology and Neurosurgery. The sample set includes two cases of vCJD, three cases of spCJD and three neurologic controls. Preliminary data using a panel of seven molecular markers is able to distinguish vCJD from spCJD using a heuristic clustering algorithm. One of the molecular markers has been identified as apolipoprotein E which appears to be upregulated in the cerebrospiral fluid (CSF) of patients with vCJD as compared to spCJD. Analysis of ante mortem CSF may help to differentiate patients with vCJD from those patients with spCJD.

Adult↗

False positive serology in cerebrospinal fluid associated with a spinal cord tumor.

Biologic false positive serology in cerebrospinal fluid has been reported as exceedingly rare. In a patient with a spinal cord tumor and elevated cerebrospinal fluid protein, the cerebrospinal fluid was reactive to VDRL and fluorescent treponemal antibody absorption (fta-abs) tests, but became nonreactive with removal of the tumor. This biologic false positive reaction may be related to the elevated cerebrospinal fluid protein, as similar false positive reactions have occurred in blood with abnormal or elevated proteins. Cerebrospinal fluid reactive to the VDRL and FTA-ABS tests does not always indicate neurosyphilis.

Cerebrospinal Fluid Proteins↗

Nitric oxide synthase is present in the cerebrospinal fluid of patients with active multiple sclerosis and is associated with increases in cerebrospinal fluid protein nitrotyrosine and S-nitrosothiols and with changes in glutathione levels.

Nitric oxide (NO) is hypothesized to play a role in the immunopathogenesis of multiple sclerosis (MS). Increased levels of NO metabolites have been found in patients with MS. Peroxynitrite, generated by the reaction of NO with superoxide at sites of inflammation, is a strong oxidant capable of damaging tissues and cells. Inducible NO synthase (iNOS) is up-regulated in the CNS of animals with experimental allergic encephalomyelitis (EAE) and in patients with MS. In this study, Western blots of cerebrospinal fluid (CSF) from patients with MS demonstrated the presence of iNOS, which was absent in CSF from control subjects. There was also NOS activity present in both MS and control CSF. Total NOS activity was increased (by 24%) in the CSF from MS patients compared with matched controls. The addition of 0.1 mM ITU (a specific iNOS inhibitor) to the samples did not change the activity of the control samples but decreased the NOS activity in the MS samples to almost control levels. The addition of 1 mM L-NMMA (a nonisoform specific NOS inhibitor), completely inhibited NOS activity in CSF from control and MS subjects. Nitrotyrosine immunostaining of CSF proteins was detectable in controls but was greatly increased in MS samples. There were also significant increases in CSF nitrate + nitrite and oxidant-enhanced luminescence in MS samples compared with controls. Additionally, a significant decrease in reduced glutathione and significant increases in oxidized glutathione and S-nitrosothiols were found in MS samples compared with controls. Parallel changes in NO metabolites were observed in the plasma of MS patients, compared with controls, and accompanied a significant increase of reduced glutathione. These data strongly support a role for nitrosative stress in the pathogenesis of MS and indicate that therapeutic strategies focussed on decreasing production of NO by iNOS and/or scavenging peroxynitrite may be useful in alleviating the neurological impairments that occur during MS relapse.

Adult↗

Identification of two-dimensionally separated human cerebrospinal fluid proteins by N-terminal sequencing, matrix-assisted laser desorption/ionization--mass spectrometry, nanoliquid chromatography-electrospray ionization-time of flight-mass spectrometry, and tandem mass spectrometry.

Optimal application of biological mass spectrometry (MS) in combination with two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) of human cerebrospinal fluid (CSF) can lead to the identification of new potential biological markers of neurological disorders. To this end, we analyzed a number of 2-D PAGE protein spots in a human CSF pool using spot co-localization, N-terminal sequencing, matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) and nanoliquid chromatography-electrospray ionization-time of flight-mass spectrometry (nanoLC-ESI-TOF-MS) with tandem MS switching. Our constructed CSF master contained 469 spots after image analysis and processing of 2-D gels. Upon visual inspection of our CSF master with the CSF pattern available on the ExPASy server, it was possible to locate and annotate 15 proteins. N-terminal sequence analysis and MALDI-MS peptide mass fingerprint analysis of both silver- and Coomassie Brilliant Blue (CBB) G-250-stained protein spots after in situ trypsin digest not only confirmed nine of the visually annotated spots but additionally resolved the identity of another 13 spots. Six of these proteins were not annotated on the 2-D ExPASy map: complement C3 alpha-chain (1321-1663), complement factor B, cystatin C, calgranulin A, hemoglobin beta-chain, and beta-2-microglobulin. It was clear that MALDI-MS identification from CBB G-250-stained, rather than from silver-stained, spots was more successful. In cases where no N-terminal sequence and/or no clear MALDI-MS result was available, nanoLC-ESI-TOF-MS and tandem MS automated switching was used to clarify and/or identify these protein spots by generating amino acid sequence tags. In addition, enrichment of the concentration of low-abundant proteins on 2-D PAGE was obtained by removal of albumin and immunoglobulins from the CSF pool using affinity chromatography. Subsequent analysis by 2-D PAGE of the fractionated CSF pool showed various new silver-stainable protein spots, of which four were identified by nanoLC-ESI-TOF-MS and tandem MS switching. No significant homology was found in either protein or DNA databases, indicating than these spots were unknown proteins.

Amino Acid Sequence↗

Proteome analysis of cerebrospinal fluid proteins in Alzheimer patients.

By comparing the CSF proteome between Alzheimer disease (AD) patients and controls it may be possible to identify proteins that play a role in the disease process and thus to study the pathogenesis of AD. We used mini-gel technology in a two-dimensional electrophoresis procedure, sensitive SYPRO Ruby staining and mass spectrometry for clinical screening of disease-influenced CSF proteins in 15 AD patients and 12 controls. The levels of six proteins and their isoforms, including proapolipoprotein, apolipoprotein E, beta-2 microglobulin, retinol-binding protein, transthyretin, and ubiquitin, were significantly altered in CSF of AD patients. The most prominently altered proteins were the apolipoproteins, especially proapolipoprotein.

Aged↗

[Cerebrospinal fluid proteins: III. Normal values of immunoglobulins G, A and M (variations related to race, sex and age)].

The literature on quantitation of immunoglobulins in normal CSF and variations of these values related to race, sex and age was reviewed. Immunoglobulins (Ig) G, A and M of normal CSF (sub-occipital puncture) of 116, 78 and 45 patients respectively, were measured by radial immunodiffusion, in order to verify variations related to race, sex and age, as well as to establish their normal limits. The results allowed us to conclude that: a) there are no differences between races with respect to the levels of IgG, A and M; b) variations related to sex or age on the CFS immunoglobulins content in children (1 to 11 years old) are not found; c) the mean IgG (mg/100 ml) level in children is lower than in adults, although the mean IgG percentage concentration in children is not different from that found in adults; the normal range for CSF IgG in children is 0.21 to 2.93 mg/100 ml; d) the mean IgA content (mg/100 ml and % of the total protein) in children are lower than in adults and in physiological conditions do not exceed 0.15 mg/100 ml (0.7% of the total protein); e) there are no differences related to sex or age with respect to the CSF levels of immunoglobulins in adults; the normal range of IgG in adults is 0.51 to 4.00 mg/100 ml; in children and adults the normal limits for the relative values of IgG are 3 to 12%; IgA is found in the CFS of adults in a concentration up to 0.32 mg/100 ml (0.9%); f) IgM is found in both children's and adults' CSF at very low levels, not exceeding 0.2 mg/100 ml (0.25%). Comparison of the results obtained in this work with those found in some publications was carried out and is briefly discussed.

Adolescent↗

Studies on protein methyltransferase in human cerebrospinal fluid.

Protein methyltransferases, rich in most mammalian brains, were studied in human cerebrospinal fluid (CSF). Among several well-characterized groups of methyltransferases, protein methylase I (S-adenosylmethionine:protein-arginine N-methyltransferase, EC 2.1.1.23) was found in significant amounts in human CSF samples. Both myelin basic protein (MBP) -specific and histone-specific protein methylase I activities were observed, the latter being generally higher in most CSF. S-Adenosyl-L-homocysteine, a potent product inhibitor for the methyltransferase, inhibited approximately 90% of MBP-specific protein methylase I activity at a concentration of 1 mM. The optimum pH of the MBP-specific protein methylase I was found to be around 7.2. Identity of exogenously added MBP as the methylated substrate for CSF enzyme was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. An amino acid analysis of the [methyl-3H]protein hydrolysate showed two major radioactive peaks cochromatographing with monomethyl- and dimethyl (symmetric)-arginine. Human CSF contained relatively high endogenous protein methylase I activity (activity measured without added substrate protein): The endogenous substrate can be immunoprecipitated by antibody raised against calf brain MBP. Finally, CSF from several neurological patients were analyzed for protein methylase I, and the results are presented.

Humans↗

Influence of methotrexate on results by three methods for determining protein in cerebrospinal fluid.

Because high intravenous and intrathecal doses of methotrexate are used in treatment for various neoplasms, this drug can reach high concentrations in cerebrospinal fluid. At those encountered clinically, it produced significant positive interference with values for cerebrospinal fluid protein as determined with the du Pont aca. A small interference was found with the Folin-Lowry method, and virtually no interference with an automated immunochemical procedure.

Cerebrospinal Fluid Proteins↗

Measurement of reference values for certain proteins in cerebrospinal fluid.

Reference values are presented for certain cerebrospinal fluid proteins and related indexes, based on determinations carried out on 46 patients with diffuse lower back pain using radial immunodiffusion and electroimmunoassay. A sex difference was noted in total protein and albumin values. The electroimmunoassay method with its sensitivity enhanced by the use of Coomassie Blue staining and optimal antiserum concentrations enabled a reference value to be determined for IgA in cerebrospinal fluid.

Adult↗

Protein determination in cerebrospinal fluid by protein dye-binding assay.

In this study, Coomassie brilliant blue (CBB) and pyrogallol red/molybdate (PRM) protein dye-binding assays for total protein determination in cerebrospinal fluid (CSF) are compared. Using human albumin (HA) as a protein calibrator, protein concentration in CSF samples (n = 73) ranged from 55-1960 mg/L (median: 315 mg/L) with the CBB assay, and from 95-2450 mg/L (median: 395 mg/L) with the PRM assay. Linear regression analysis indicated yCBB = 0.824xPRM - 8 (r = 0.99). The discrepancy between the values was investigated by comparing the response of the two assays to different proteins. Compared with HA, the PRM assay showed a more uniform response to human albumin/globulin (A/G) and bovine gamma globulin (G) than did the CBB assay, but it gave high colour yields with bovine myelin basic protein. When CSF was assayed using A/G as a protein calibrator, agreement between the methods improved (yCBB = 0.960xPRM + 0 [r = 0.99]), indicating that comparability is dictated by the choice of protein calibrator. Of the two assays studied, the PRM assay is recommended for CSF protein determination because it gives a more uniform and linear response to human albumin and globulin over a wider working range.

Animals↗

[Effect of protein-containing cerebrospinal fluid on ventriculo-auricular drainage].

Elevated protein level in liquor still prohibits ventriculo-auricular drainage methods. Spitz-Holter valves and Pudenz-Heyer valves were therefore tried in model tests using plasma-containing fluids. The quantities of flow depending on time were presented on graphs. A considerably reduced flow through the Spitz-Holter valve was observed after brief perfusion already, with tendency to complete occlusion. Similar results were obtained from trials on the Pudenz-Heyer valve.

Cerebrospinal Fluid Proteins↗