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PubMed2003

Improved resolution of human cerebrospinal fluid proteins on two-dimensional gels.

Proteomics combines two-dimensional gel electrophoresis and peptide mass fingerprinting and can potentially identify a protein(s) unique to disease. Such proteins can be used either for diagnosis or may be relevant to the pathogenesis of disease. Because patients with multiple sclerosis (MS) have increased amounts of immunoglobulin (Ig) G in their cerebrospinal fluid (CSF) that is directed against an as yet unidentified protein, we are applying proteomics to MS CSF, studies that require optimal separation of proteins in human CSF. We found that recovery of proteins from CSF of MS patients was improved using ultrafiltration, rather than dialysis, for desalting. Resolution of these proteins was enhanced by acetone precipitation of desalted CSF before electrophoresis and by fractionation of CSF using Cibacron Blue sepharose affinity chromatography. Improved protein recovery and resolution will facilitate excision from gels for analysis by peptide mass fingerprinting.

Albumins↗
PubMed1976

[Composition of cerebrospinal fluid protein in children with nervous system diseases according to disc-electrophoresis in polyacrylamide gel].

The author studied the fractional content of proteins in the CSF in 105 children with different disorders of the nervous system and 35 children of a control group (aged 2-13 years) with the aid of disc-electrophoresis in polyacrylamide gel. The proteins of the CSF were studied in encephalitis, polyradiculoneuritis, cerebral arachnoiditis, brain tumors and epilepsy. The content of total proteins in the CSF in 42% of the studied children did not differ significantly from the usual age norm. More distinct changes were found in proteinogramms of the CSF where 14 protein fractions were singled out. It was established that a change in the content of separate fractions, their intercorrelation depended on the character of the pathological process.

Adolescent↗
PubMed1975

[The relationship between age and the cerebrospinal fluid protein profile profile of normal children. Cellulose acetate and agarose gel electrophoretic studies].

Neuropädiatrie 6: 383--397 (1975). CSF samples of 63 normal children 2 days to 14 years of age were concentrated (ultrafiltration in "Kollodiumhülsen") and examined by electrophoresis on cellulose acetate and agarose gel. In the CSF of all children 7 protein fractions were found by cellulose acetate electrophoresis. Analysing the pherograms by the means of an analog computer 15 fractions could be separated by agarose gel electrophoresis. Compared with the children of school age the newborn showed low prealbumin, beta1-globulin and tau-globulin fractions besides high albumin and gamma-globulin concentrations owing to an increased permeability of the blood-CSF barrier. During the following weeks a low prealbumin and a high albumin fraction still was found. Children from 3 month to 2 years of age showed a moderate increase of the relative prealbumin and beta1-globulin concentration in comparison with older children. The gamma-globulin fraction was lowest beyond the third month of life and increased again in the group of the 2--14 years old children. The variation of the protein profile with age could not only be demonstrated by the more difficult agarose gel electrophoresis but also by a simple technique of cellulose acetate electrophoresis.

Adolescent↗
PubMedDate not supplied

Cerebrospinal fluid proteins electrophoresis without prior concentration.

There is a need for new methods to study CSF proteins. Kerenyi et al., 1973 and Verheecke, 1975 have already used this method and discussed its advantages. The electrophoresis is carried out according to the method of Wieme with as supporting medium agar. The volume applied varies between 5 to 15 microliters according to the total protein content and the width of the slit. Care must be taken to work at all times with bidistilled water and pro analysis reagents. After drying, the electrophoretic plates are reduced in a 10% solution of thiodiglycol, dried again, then immersed in 2% potassium hexacyanoferrate, washed thoroughly, then revealed with a silver nitrate solution (Kerenyi et al., 1973; Verheecke P., 1975) and left in 1% acetic acid. This technique is of major value as: 1. It avoids artefacts due to concentration and loss of proteins; 2. It works with very small amounts of fluid; 3. Where the CSF is silent for antibodies restricted heterogeneity using the classical methods, it reveals marked IgG fractionation.

Cerebrospinal Fluid Proteins↗
PubMedDate not supplied

Cerebrospinal fluid protein electrophoresis without prior concentration.

There is a need for new methods to study CSF proteins. Kerenyi et al., 1973 and Verheecke, 1975 have already used this method and discussed its advantages. The electrophoresis is carried out according to the method of Wieme with as supporting medium agar. The volume applied varies between 5 to 15 microliter according to the total protein content and the width of the slit. Care must be taken to work at all times with bidistilled water and pro analysis reagents. After drying, the electrophoretic plates are reduced in a 10% solution of thiodiglycol, dried again, then immersed in 2% potassium hexacyanoferrate, washed thoroughly, then revealed with a silver nitrate solution (Kerenyi et al., 1973; Verheecke P., 1975) and left in 1% acetic acid. This technique is of major value as: 1. It avoids artefacts due to concentration and loss of proteins; 2. It works with very small amounts of fluid; 3. Where the CSF is silent for antibodies restricted heterogeneity using the classical methods, it reveals marked IgG Fractionation.

Animals↗
PubMed1983

[Valve of the use of benzethonium chloride for turbidimetric analysis of cerebrospinal fluid proteins with an automatic analyzer].

We have adapted Iwata and Nishikaze's benzethonium chloride turbidimetric method for the assay of CSF proteins to the techniques of continuous flow and the centrifuge analyser. The proposed technique presents several advantages: benzethonium chloride forms a stable precipitate with the proteins with a slow kinetic of formation, enabling adaptation to the centrifuge analyser; the response obtained is identical for albumin and immunoglobulin G; this very sensitive technique allows the use of micro-samples. Studies of repeatability and the correlations made with other techniques give very satisfactory results.

Autoanalysis↗
PubMed1988

Cerebrospinal fluid protein analysis in diseases of the nervous system.

Electrophoretic studies of human CSF proteins from patients with diseases of the NS are reviewed. Various 1-DE methods are of similar value in identifying the non-specific OBs, which are helpful in the diagnosis of MS and recurrent GBS. In early and subclinical MS, OBs are of prognostic value, with IEF gels having the greater resolution. Silver-stained 2-DE gels provide the equivalent information to the OBs on 1-DE gels, with even greater sensitivity, and yield additional disease-associated protein data. Two proteins have proven to have diagnostic value in CJD and other changes that are still being evaluated have been identified in Parkinson's disease, GBS, Alzheimer's disease, schizophrenia and Herpes simplex encephalitis. The vastly improved CSF protein information obtained with silver-stained 2-DE gels heralds both a change from the relatively limited applications with 1-DE methods and also the need to adopt this approach in the routine clinical laboratory.

Animals↗
PubMedDate not supplied

[Evaluation of cerebrospinal fluid proteins in patients with subacute sclerosing panencephalitis (SSPE)].

CSF proteins of 62 patients with SSPE aged 6 to 26 years were analysed by polyacrylamide gel electrophoresis. The total IgG concentration in the CSF was increased to 16.8 +/- 10 mg/dl and the IgG index was raised to from 2.7 +/- 0.5 to 3.6 +/- 3.0. All SSPE patients had pathological subfractions gamma 4 and gamma 5. The per cent proportion of the gamma 1 subfraction was raised only in the youngest patients. A tendency was observed for higher values of the prealbumin:albumin index and the prealbumin: transferrin index. The youngest SSPE patients had a tendency for greater total protein concentration, lower prealbumin concentration, and highest IgG index values (3.6 +/- 3.0). In early disease period a tendency was noted for higher concentration of total protein, higher total IgG level and higher IgG index. These results suggest that SSPE patients have a constant stimulation of B-cells and variable increase of blood-brain barrier permeability.

Adolescent↗
PubMed1975

Analytical isotachophoresis: a new method for analysis of cerebrospinal fluid proteins. A preliminary report.

Isotachophoresis of CSF proteins seems to be a very promising method. Very small CSF samples, a few mul of concentrated and 15-30 mul of unconcentrated CSF, can be quickly analysed (30-60 min), and the results immediately obtained on a recorder. Using unconcentrated CSF, losses due to concentration procedures, are avoided. Low-molecular weight compounds, e.g. in CSF ultrafiltrates, can also be examined. The method gives high resolution, is reproducible, and is easy to perform. The isotachophoretic findings have been compared with those of electrofocusing.

Cerebrospinal Fluid Proteins↗
PubMed1987

Serum and cerebrospinal fluid proteins and the blood-brain barrier in Alzheimer's disease and multi-infarct dementia.

Serum concentrations of IgG, IgA, IgM, haptoglobin, transferrin, prealbumin and albumin quantitated nephelometrically in 22 patients with Alzheimer's disease (AD), 29 patients with multi-infarct dementia (MID) and their age-matched controls were normal. Cerebrospinal fluid (CSF) albumin and CSF/serum ratio for albumin were higher in AD and MID patients compared to controls, but no significant differences were found between AD and MID. Patients with MID had elevated CSF IgG, IgA, IgM and prealbumin levels compared to controls and to AD. An increased CSF IgG index was found in 5 MID patients but none of the AD patients. Thus, the blood-brain barrier permeability is often increased in MID as well as in AD. There is no increased intrathecal IgG synthesis in AD but it may occur in MID.

Aged↗
PubMed1991

Age-related changes in cerebrospinal fluid protein concentrations.

The blood-brain barrier (BBB) has a key role in CSF homeostasis and preservation of normal neuronal function. There has been little investigation of changes in barrier function in elderly subjects without evidence of neurological disease. The ageing brain demonstrates increased vulnerability to a variety of insults, which may reflect a deterioration in barrier performance. Direct measurement of BBB function is difficult but the CSF/serum quotients of individual proteins is the most widely used parameter. The present study sought age related changes in the CSF/serum quotients of prealbumin, albumin, immunoglobulin G and alpha 2-macroglobulin in adults with no neurological deficit. A prospective study was established, and 107 subjects (aged 18-89 yrs) were recruited from patients undergoing spinal anaesthesia or diagnostic myelography. A weak but significant positive correlation was found between the CSF/serum quotients and age for all the proteins studied, but with the exception of alpha 2-macroglobulin, these changes probably reflect age related falls in serum protein concentrations found in this study. alpha 2-Macroglobulin appears to be a sensitive indicator of barrier function for large populations, suggesting a subtle decline of BBB integrity with increasing age. This may be a true age-related change or could reflect an increased incidence of subclinical disease in an ageing population.

Adolescent↗
PubMedDate not supplied

[Intrathecal synthesis of immunoglobulins during acute viral meningitis with high cerebrospinal fluid protein levels].

The article deals with the results of examination of immunoglobulins (IgM, IgA, IgG) in the liquor and serum of 50 patients with acute viral meningitis and hyperproterinorashia over 1.0 g/l. The quantitative determination of immunoglobulins was done by the method of radial immunodiffusion according to Manzoni. The confirmation of intrathecal synthesis of immunoglobulins was done by liquor indexes. IGM intrathecal synthesis was found in 11 (22%) patients, IgA in 16 (32%) and IgG in 10 (20%) subjects. The daily amount of IgG synthesis in the central nervous system was 1.65-97.15 mg (mean 26.60 mg). The values of liquor indexes were decreased in later disease. This suggested the decrease of disappearance of immunoglobulin intrathecal synthesis. There were 26 control patients with meningism (meningeal syndrome with regular cytobiochemical finding in the liquor). IgG intrathecal synthesis was established in one patient with meningism. This finding was an extremely rare and unusual finding.

Acute Disease↗
PubMed2004

Meningeal derived cerebrospinal fluid proteins in different forms of dementia: is a meningopathy involved in normal pressure hydrocephalus?

OBJECTIVES: In animal models and in vitro studies leptomeninges have been shown to be the origin of neurotrophic substances that support the survival and growth of neuronal cells. Because dementia is associated with neuronal loss, we investigated whether leptomeningeal dysfunction may be involved in the pathogenesis of dementia disorders. METHODS: We analysed the cerebrospinal fluid (CSF) concentrations of the leptomeningeal derived beta trace protein, beta2 microglobulin, and cystatin C. RESULTS: There was a statistically significant difference of the CSF beta trace protein levels among different groups. Patients with idiopathic normal pressure hydrocephalus (NPH) (17.5 (SD 4.3) mg/l) showed significantly lower CSF beta trace protein levels than patients with Alzheimer's disease (23.8 (6.2) mg/l), depression (24.2 (7.3) mg/l), and normal controls (25.3 (4.9) mg/l). To patients with vascular dementia (20.1 (5.6) mg/l) and frontotemporal dementia (21.9 (7.0) mg/l), the difference was not significant. There was no significant difference regarding the CSF and serum concentrations of beta2 microglobulin or cystatin C among the different groups. CONCLUSIONS: We conclude that leptomeningeal dysfunction may be involved in certain types of dementia such as NPH and that reduced CSF beta trace protein levels in patients with NPH may aid in differentiating this difficult to diagnose disorder from other syndromes such as Alzheimer's disease.

Aged↗
PubMed1975

Cerebrospinal fluid proteins in multiple sclerosis.

Various CSF proteins were studied in 255 definite multiple sclerosis patients at various disease stages and compared with corresponding values obtained from 174 controls. The CSF changes in acute multiple sclerosis patients included a significant increase of total proteins and of gamma globulin, IgG, IgA, IgM, alpha-2 ceruloplasmin, 7S-gamma-1, and cytotoxic index for nerve cells in tissue culture, and significant decreases of pre-albumin, alpha-1, and alpha-2 and of the beta/gamma globulin ratio. The CSF levels of IgG, IgA, and IgM remained significantly higher in steroid-treated multiple sclerosis patients than in controls, but the levels often were significantly reduced while patients were on treatment or in remission. During remission or treatment with ACTH and/or steroids, the alpha-2 ceruloplasmin, 7S-gamma-1, and cytotoxic index were significantly reduced and the pre-albumin, alpha-1, and alpha-2 globulin classes and the beta/gamma ratio showed a tendency to return to normal.

Adolescent↗
PubMed1976

Isoelectric focusing and electrophoresis of cerebrospinal fluid proteins in muscular dystrophies and spinal muscular atrophies.

In the very few previous investigations of the CSF-proteins in muscular dystrophies the results have generally been reported as normal. In spinal muscular atrophies a barrier-damage pattern of CSF-proteins has been found in amyotrophic lateral sclerosis (ALS). In the present investigation the CSF-proteins were examined by isoelectric focusing and quantitative paper electrophoresis in 13 patients with muscular dystrophies and in 11 patients with spinal muscular atrophies. On isoelectric focusing, CSF-protein abnormalities were found in 85% of the cases with muscular dystrophies and in all patients with spinal muscular atrophies. Differences in the CSF-protein patterns were observed within the group of muscular dystrophies and between these and the cases of spinal muscular atrophies. In ALS and in myotonic dystrophy, abnormal CSF-protein fractions occurred mainly in the alkaline pH-range, while in limb-girdle dystrophy and the patient with facioscapulohumeral dystrophy, aberrant fractions appeared mainly in the acidic region. CSF-protein abnormalities were found in both the alkaline fractions (HAFs) with pI 9.2-9.6 and a fraction with PI 7.1 were found in half of the patients with myotonic dystrophy. The CSF electrophoresis in myotonic dystrophy showed increased levels of beta1-globulin in all cases examined. Signs of barrier-damage were commonly encountered in ALS in contrast to the muscular dystrophies, except for myotonic dystrophy. The results are discussed in terms of possible diagnostic value and with regard to pathogenetic significance, particularly in relation to the current hypothesis of a neural involvement in muscular disorders.

Adult↗
PubMedDate not supplied

[Electrophoresis of cerebrospinal fluid proteins in patients with ischemic cerebrovascular disease].

INTRODUCTION: Cerebral infarction (CI) increases vascular permeability because of a torrent of molecular events that take place. It frequently leads to oedema, haemorrhage and neuronal death. Free radicals and proteases are also formed, which cause lesions in the blood vessels, and microvascular integrity is lost through degradation of the basal lamina and the extracellular matrix. As a result rupture of the blood brain barrier takes place. AIMS. To compare the electrophoretograms of patients with ischemic cerebrovascular disease (ICVD) with those of controls and to link the alterations in the proteinogram with the ICVD subtypes. PATIENTS AND METHODS: The CSF of 55 controls and 136 patients with ICVD was examined. The total protein (TP) concentration was determined and a polyacrylamide gel electrophoretogram was produced using Coomassie blue stain. Parallel to this, serum was prepared for haptoglobin staining. RESULTS: The TP in patients CSF rose to a significantly higher level than that of controls. Blood brain barrier damage (BBBD) was observed in 28.7% of the patients studied with CI and in 10.3% in transient ischemic attacks (TIA), while 16.2% presented oligoclonal bands. There was a difference between the two sexes: men were found to have higher TP levels, lower percentages of prealbumin 1 and more BBBD than women. CONCLUSIONS: BBBD is more frequent in infarctions than in TIA, and is predominant in men with thrombotic cerebral infarction.

Adult↗