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Standardization of the Coomassie Blue method for cerebrospinal fluid proteins.

Coomassie Brilliant Blue G-250 can be used to quantitatively determine proteins in cerebrospinal fluid. When the dye combines with protein, the absorption maximum of the dye shifts. The dye-protein color forms almost instantaneously and is stable for at least 1 h. The procedure is also insensitive to changes in temperature in the range of 20--30 degrees C. The absorptivities of the dye complexes with human albumin or globulin differ, thus a pure albumin or pure globulin standard is unsuitable; a standard containing 70% albumin and 30% globulins is the most appropriate for this application. A bichromatic approach to standardization increases the range of linearity of a calibration curve. The method gives values that are about 9% higher than a sodium sulfate-sulfosalicylic turbidimetric procedure for cerbrospinal fluid proteins.

Cerebrospinal Fluid Proteins

[Changes in cerebrospinal fluid protein concentration in children following surgery for cerebellar medulloblastoma].

In 10 children after operations for cerebellar medulloblastoma treated with intrathecal injections of methotrexate changes were studied in the concentration of total protein in the cerebrospinal fluid and in different electrophoretic fractions of proteins. The investigations were begun 1 to 3 months after the operation and were continued usually at monthly time intervals. In each child considerable variations were observed in the concentration of total protein as well as in the proportions of protein electrophoretic fractions in the period of treatment lasting from 14 to 59 months after the operation. Significant individual variations were observed also in the level of total protein fractions pattern in the cerebrospinal fluid. Out of 116 electrophorograms in 42 the protein fraction pattern in the cerebrospinal fluid approached very much that found in patients with cerebellar medulloblastoma before the operation, with a significant rise in the proportion of albumins. The origin of cerebrospinal fluid proteins is discussed.

Cerebellar Neoplasms

[Behavior of cerebrospinal fluid proteins in degenerative diseases].

132 cerebrospinals fluids from patients with degenerative diseases of the central nervous system have been analyzed for protein distribution in agar gel electrophoresis. After subdivision into diagnostically well defined groups these patients were compared with 48 with metabolic and psychiatric diseases and with 79 normal controls. The majority of diagnostic groups showed a tendency to permeability impairment. Other outstanding deviations were not found, except for single cases which were not statistically typical of the groups as a whole. However, the "degenerative type" of proteinogram emphasized in the literature predominated not only in the degenerative groups but also in certain other diseases with destruction of central nervous tissue which do not belong to the degenerative diseases in the strict sense. On the other hand, there are some degenerative subgroups without this "typical" electrophoretic pattern. There would thus appear to be grounds for amending the term "degenerative" into "tissue destroying" or "atrophic type", to avoid misinterpretation.

Alpha-Globulins

Electrophoresis of normal lumbar cerebrospinal fluid proteins in the African.

Lumbar cerebrospinal fluid obtained from Zambian patients suspected of suffering from central nervous system conditions was concentrated over a hundred-fold and subjected to small-scale cellulose acetate electrophoresis (Millipore). The percentage of each protein fraction was determined by scanning. Only twenty samples were considered normal out of a total of 150 studies. These normal values were not statistically different from those of published data on Europeans and North American Caucasians. The high serum gamma globulin in African subjects is not reflected in the cerebrospinal fluid.

Adolescent

Hormones, electrolytes, and cerebrospinal fluid proteins in manic-melancholic patients.

The concentrations of insulin and thyroid hormones, tryptophan, electrolytes, urea, plasma proteins in the cerebrospinal fluid, and glucose in blood and cerebrospinal fluid in manic-melancholic patients were studied. As control groups served patients suffering from other psychiatric disorders as well as neurological and orthopedic patients. Apart from the blood values of thyroid hormones, the results showed no differences between the various diagnostic groups, neither in the abnormal states nor when recovered. For blood thyroxine and free thyroxine index, a statistically significant differences was seen in unipolar (melancholic) patients, namely a decrease concomitant with the clinical improvement. A tendency in the opposite direction of the thyroxine values was found in bipolar (melancholic) patients. In the manic group a marked decrease in the thyroxine values was obtained in the lithium-treated patients.

Adolescent

Cerebrospinal fluid protein findings in various lower back pain syndromes.

Cerebrospinal fluid (CSF) total protein, albumin and IgG concentrations were measured in 53 patients with lower back pain syndromes. In the majority of the patients (81 %) the protein values were within normal ranges, a finding contrary to previous studies. In six of 28 patients with acute lumbar disc prolapse and in two of four cases with spinal stenosis, a clearly abnormal protein pattern was observed, while patients who had been earlier operated on for disc prolapse showed normal values. The calculated permeability indexes suggest that the elevation of various proteins is linked with increased permeability across the blood-brain barrier and that the local immunoglobulin synthesis in CNS is lacking. In only one of the cases with an abnormal CSF protein pattern could no evidence of the cause of the pathological finding be observed in subsequent examinations.

Adult

The normal cerebrospinal fluid proteins identified by means of thin-layer isoelectric focusing and crossed immunoelectrofocusing.

The cerebrospinal fluid and serum proteins from 32 healthy individuals have been examined by isoelectric focusing in a pH-gradient from 3.5--11.0. Seventeen normal proteins have been identified by means of crossed immunoelectrofocusing. The pI-values of the main fractions of these proteins have been determined. On crossed immunoelectrofocusing microheterogeneity was observed for all of the proteins except haemopexin, gamma-trace protein of CSF and prealbumin of serum. Differences between serum and CSF were observed for 6 of the proteins. Prealbumin of CSF had a lower pI-value than in serum and showed partial immunological identity with prealbumin of serum. Transferrin of CSF included several components with lower sialic acid contents than in serum. The conversion of C'3-complement from beta-1-C to beta 1-A was slower in CSF than in serum. Qualitative differences between CSF and serum were also noticed for alpha2-macroglobulin and haptoglobin. IgM was not detected in CSF. In CSF the gamma-trace protein was found on isoelectric focusing in one third of the subjects. Two subjects had 2 close tau-fractions and another two had one unidentified band in the anodal IgG-area. The gamma-trace protein did not show any proteolytic activity, nor any resemblance in charge to 4 basic enzymes or with the histones tested.

Adult

Agarose gel electrophoresis of cerebrospinal fluid proteins and analysis of the pherogram profiles by analog computer.

A micro-method of agarose gel electrophoresis requiring 1--3 ml of cerebrospinal fluid for quantitative analysis of cerebrospinal fluid proteins is described. After concentration of CSF to about 50 microliter by ultrafiltration and refractometric determination of protein, approximately 20--40 microgram of total protein are used for electrophoresis. Photometric scanning of the electrophoretic pattern at two different wavelengths permits quantitative evaluation. The pherograms are analysed by means of a modified DU-PONT analog computer. Factors which influence quantitative electrophoresis are examined. In cerebrospinal fluid of normal children 15 protein fractions are demonstrated: 2 prealbumins, albumin, 5 alpha-, 3 beta- and 4 gamma-globulins.

Adolescent

Electrophoretic pattern of cerebrospinal fluid proteins in non-neoplastic infantile hydrocephalus.

Lumbar cerebrospinal fluid (CSF) from 28 non-neoplastic hydrocephalic children was studied for total protein and electrophoretic protein patterns. These were classified as normal, degenerative, transudative and gamma-globulinic according to Laterre. We found higher total CSF protein mean values than in normal cases with the same age and abnormal electrophoretic patterns in 72% of the cases, of which degenerative was the most common (54%). Gamma-globulinic and transudative patterns were found in 11% and 7% of the cases, respectively. Several factors which may explain the increase in the total CSF protein in infantile hydrocephalus are described: low age, ventricular block in the noncommunicating hydrocephalus, and probable passage of tissue proteins to the CSF from the damaged brain. The predominance of degenerative patterns may be explained by the enrichment of the CSF in tissue proteins resulting from the white matter damage provoked by the abnormal conditions of production, flow and absorption of the CSF in hydrocephalus. Ventricular CSF was studied in four cases, and the results obtained are in agreement with the above-mentioned findings.

Albumins

Crossed immunoelectrofocusing for identification of normal and abnormal cerebrospinal fluid proteins. A preliminary report.

Isoelectric focusing is a valuable method for the analysis of cerebrospinal fluid (CSF) proteins. Its high resolving power has, however, created problems in identifying the large number of protein bands separated. In an attempt to identify these bands, two-dimensional crossed immunoelectrofocusing has been used, where isoelectric focusing is combined with rocket immunoelectrophoresis. By these methods 9 of the normal proteins in the acidic pH interval have been identified. In addition the unusual CSF protein abnormalities occurring in Marie-Sanger-Brown's ataxia and alcoholic cerebellar degeneration have been shown to represent increases of different microheterogeneous forms of transferrin.

Alcoholism

Familial amentia, unusual ventricular calcifications, and increased cerebrospinal fluid protein.

A sibship originally reported by Friedman and Roy as showing severe mental retardation, strabismus, hyperactive tendon reflexes, lalling speech, and foot deformities was restudied. Three major additional findings were noted. The cerebrospinal fluid protein concentration was increased two to three times above normal in four siblings who were available for study. Radiographs of cranial structures in three siblings showed identical pathologic intracranial calcifications which correspond in distribution to the choroid plexus. The choroid plexus was not demonstrable in one patient when radiolabeled 99m-Tc-pertechnetate was injected without perchlorate. Neuropathologic findings in one sibling included small subcortical heterotopias and atrophy of the choroid plexus with encasement by glial fibrils. These findings denote a new heredofamilial neurologic syndrome associated with mental retardation and a disorder of choroid plexus.

Atrophy

[Value of cerebrospinal fluid protein study by the polyacrylamide gel electrophoresis method].

The methods of cerebrospinal fluid electrophoresis used as yet are surveyed briefly with particular reference to the method using polyacrylamide gel. In the light of own results it has been found that the method is superior because only small amounts of material are required and the sensitivity and separating ability of the method are much higher.

Cerebrospinal Fluid Proteins

[Cerebrospinal fluid protein electrophoresis in non-inflammatory central nervous system diseases].

Cerebrospinal fluids (CSF) (N = 365) from patients with non-inflammatory diseases of the central nervous system were analyzed for protein distribution by agar gel microelectrophoresis. After subdivision into diagnostically well defined groups, these patients were compared with 79 normal controls. Most of the diseases investigated were found to follow the "plasma-type" pattern. In some of them the deviations from normal were so extensive that a CSF-protein pattern similar to the "acute-phase reaction" in the serum occurred. Moreover, the following characteristics were found: a considerable increase in total protein and the gamma-globulin fractions in neurinomas; a reproducible increase in the alpha1-globulins in metastases of the central nervous system; and a statistically significant difference of CSF protein findings between male and female patients with protrusion of lumbar intervertebral discs.

Albumins

[Bidimensional electrophoresis in the study of cerebrospinal fluid protein (author's transl)].

The authors report 650 bidimensional electrophoreses of the cerebrospinal fluid (CSF). All examinations were carried out after prior concentration of CSF protein. The technic used is that described by Rebeyrotte in 1970. Four types of tracing were noted: -- Two tracings were unusual by the presence of a large peak in the alpha-2 region. -- One tracing was of the hypergamma type. -- One tracing was comparable to serum by the richness of the precipitations. A change in the first migration permitted us to obtain electrophoretic separations from 100 microlitres of pure CSF. The bidimensional immunoelectrophoresis tracings are comparable to those described previously.

Cerebrospinal Fluid Proteins