Cerebrospinal Fluid Protein Fractions Research Articles

Acute phase proteins in serum and cerebrospinal fluid in healthy cattle: possible use for assessment of neurological diseases

ABSTRACT: Use of acute-phase proteins (APPs) for assessment of health and disease in animals has increased greatly within the last decade. The objective was to determine the normal concentration of APPs in the serum and cerebrospinal fluid (CSF) of healthy cattle by polyacrylamide gel electrophoresis. Fifty crossbred animals (350±70kg of BW and 18±1.2 months of age), 25 heifers and 25 steers were used. CSF samples were collected from atlanto-occipital (AO) site and blood samples were obtained from the jugular vein. CSF and serum protein electrophoresis were performed by means of sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Thirty-seven proteins with molecular weights ranging from 7 and 37kDa were identified in CSF of all animals. These eight were nominally identified with immunoglobulin A and G, celuloplasmin, transferrin, albumin, α1-antitripsin, acidic glycoprotein, and haptoglobin. All protein fractions in CSF did not differ between heifers and steers. In sera, 34 proteins with molecular weights between 7 and 244kDa were identified in heifers and steers. Similar proteins were nominally identified in the sera, but only the CSF presented α1-antitripsin. The serum values of acidic glycoprotein and immunoglobulin G were significantly higher in steers compared with heifers. In conclusion, measurement of CSF acute phase protein concentrations can be useful in diagnosing and monitoring the progression of bovine neurological diseases, perhaps even to guide therapeutic procedures. The CSF electrophoretic profile of healthy cattle does not change depending on gender.

Most of the Abundant Protein Fractions of Embryonic Cerebrospinal Fluid are Produced Out of the Brain Anlagen

The microenvironment of the central nervous system is important for neuronal function and development. Dur- ing the early stages of embryo development the cephalic vesicles are filled by embryonic cerebrospinal fluid, a complex fluid containing different protein fractions, which contributes to the regulation of the survival, proliferation and neuro- genesis of neuroectodermal stem cells. The protein content of embryonic cerebrospinal fluid from chick and rat embryos at the start of neurogenesis has already been determined. Most of the identified gene products are thought to be involved in the regulation of developmental processes during embryogenesis. However, due to the crucial roles played by embry- onic cerebrospinal fluid during brain development, the embryological origin of the gene products it contains remains an intriguing question. According to the literature most of these products are synthesised in embryonic tissues other than the neuroepithelium. In this study we examined the embryological origin of the most abundant embryonic cerebrospinal fluid protein fractions by means of slot-blot analysis and by using several different embryonic and extraembryonic protein ex- tracts, immunodetected with polyclonal antibodies. This first attempt to elucidate their origin is not based on the proteins identified by proteomic methods, but rather on crude protein fractions detected by SDS-PAGE analysis and to which polyclonal antibodies were specifically generated. Despite some of the limitations of this study, i.e. that one protein frac- tion may contain more than one gene product, and that a specific gene product may be contained in different protein frac- tions depending on post-translational modifications, our results show that most of the analysed protein fractions are not produced by the cephalic neuroectoderm but are rather stored in the egg reservoir; furthermore, few are produced by em- bryo tissues, thus indicating that they must be transported from their production or storage sites to the cephalic cavities, most probably via embryonic serum. These results raise the question as to whether the transfer of proteins from these two embryo compartments is regulated at this early developmental stage.

Agarose gel electrophoresis of cerebrospinal fluid proteins of dogs after sample concentration using a membrane microconcentrator technique

Cerebrospinal fluid (CSF) is produced in the cerebral ventricles through ultrafiltration of plasma and active transport mechanisms. Evaluation of proteins in CSF may provide important information about the production of immunoglobulins within the central nervous system as well as possible disturbances in the blood-brain barrier. The objective of this study was to measure the concentration and fractions of protein in CSF samples using a membrane microconcentrator technique followed by electrophoresis, and to compare the protein fractions obtained with those in serum. CSF samples from 3 healthy dogs and 3 dogs with canine distemper virus infection were concentrated using a membrane microconcentrator having a 0.5 to 30,000 d nominal molecular weight limit (Ultrafree, Millipore, Billerica, MA, USA). Protein concentration was determined before and after concentration. Agarose gel electrophoresis was done on concentrated CSF samples, serum, and serial dilutions of one of the CSF samples. Electrophoretic bands were clearly identified in densitometer tracings in CSF samples with protein concentrations as low as 1.3 g/dL. The higher CSF protein concentration in dogs with distemper was mainly the result of increased albumin concentration. The microconcentrating method used in this study enables characterization of the main protein fractions in CSF by routine electrophoresis and may be useful for interpreting the underlying cause of changes in CSF protein concentrations.

Critical evaluation of the biological role of IgM in cerebrospinal fluid in inflammatory and other diseases of the nervous system

In a group of 10,156 patients with neurological diseases, the IgM level was assessed (using laser nephelometry) both in cerebrospinal fluid (CSF) and serum; concentration of other 17 protein fractions was also simultaneously determined: albumin, immunoglobulins, acute phase reactants, complement components, apolipoproteins, proteinase inhibitors and alpha1-microglobulin. Total protein, element counts and glucose level were also evaluated. In patients with normal CSF findings, only limited statistically significant correlations were demonstrated between IgM and other CSF protein fractions while, in the group of patients with pathological CSF findings, significant correlations were found between CSF(IgM) and other immunoglobulins, complement fractions and the rate of intrathecal synthesis of immunoglobulins. Correlations were also found between CSF(IgM) and CSF antithrombin-III and alpha1-microglobulin. Correlations between CSF(IgM) and CSF apolipoproteins support the theory of CNS tissue destruction whenever the concentration of CSF apolipoproteins is elevated. Our data substantially contribute to establishing the diagnosis in patients with neurological diseases; simultaneous measurement of a high number of CSF proteins is becoming inevitable for a reasonable assessment of the CSF Protein Status.

Analysis of cerebro‐spinal fluid protein composition in early developmental stages in chick embryos

Foetal cerebro-spinal fluid (CSF) has a very high protein concentration when compared to adult CSF, and in many species five major protein fractions have been described. However, the protein concentration and composition in CSF during early developmental stages remains largely unknown. Our results show that in the earliest stages (18 to 30 H.H.) of chick development there is a progressive increase in CSF protein concentration until foetal values are attained. In addition, by performing electrophoretic separation and high-sensitivity silver staining, we were able to identify a total of 21 different protein fractions in the chick embryo CSF. In accordance with the developmental pattern of their concentration, these can be classified as follows: A: high-concentration fractions which corresponded with the ones described in foetal CSF by other authors; B: low-concentration fractions which remained stable throughout the period studied; C: low-concentration fractions which show changes during this period. The evolution and molecular weight of the latter group suggest the possibility of an important biological role. Our data demonstrate that all the CSF protein fractions are present in embryonic serum; this could mean that the specific transport mechanisms in neuroepithelial cells described in the foetal period evolve in very early stages of development. In conclusion, this paper offers an accurate study of the protein composition of chick embryonic CSF, which will help the understanding of the influences on neuroepithelial stem cells during development and, as a result, the appropriate conditions for the in vitro study of embryonic/foetal nervous tissue cells.

Inflammatory and autoimmune diseases of the nervous system; possibilities of laboratory diagnostic methods in cerebrospinal fluid.

Contemporary aspects of cerebrospinal fluid analysis are discussed, including the relationship to neuro-infective, autoimmune and other neurological diseases. The actual state of cerebrospinal fluid microbiological and cytological investigation and analysis of cerebrospinal fluid protein fractions are described in detail.

Flow rate of cerebrospinal fluid (CSF) — A concept common to normal blood-CSF barrier function and to dysfunction in neurological diseases

Many neurological diseases are accompanied by increased protein concentrations in the cerebrospinal fluid (CSF), described as a blood-CSF barrier dysfunction. The earlier interpretation as a “leakage” of the blood-CSF barrier for serum proteins could be revised by introduction of a “population variation coefficient” of the CSF / serum quotients for IgG, IgA and IgM (ΔQ/Q̄) which is evaluated as a function of increasing albumin quotients (Q Alb). The data presented here are based on specimens from 4380 neurological patients. These population variation coefficients were found to be constant over two orders of magnitude of normal and pathological CSF protein concentrations (Q Alb = 1.6 · 10 −3 - 150 · 10 −3). This constancy indicates that there was no change in blood-CSF barrier related structures with respect to diffusion controlled protein transfer from blood into CSF and hence no change in molecular size dependent selectivity. The pathological increase of plasma protein concentrations in CSF in neurological diseases could also be explained quantitatively by a decrease of CSF flow rate due to its bifunctional influence on CSF protein concentration: reduced volume exchange, and as newly stated, increased molecular net flux into CSF without change of permeability coefficients. Again, on the basis of a changing CSF flow rate, the hyperbolic functions, which describe empirically the changing quotient ratios between proteins of different size (e.g. Q IgG : Q Alb) with increasing CSF protein content (Q Alb) can likewise be derived from the laws of diffusion as the physiologically relevant description. The hyperbolic discrimination line between brain-derived and blood-derived protein fractions in CSF in the quotient diagrams for CSF diagnosis can be further improved on the basis of the large number of cases investigated. Other physiological and pathological aspects, such as high CSF protein values in the normal newborn, in spinal blockade, in meningeal inflammatory processes, CNS leukemia or polyradiculitis as well as animal species dependent variations can each be interpreted as due to a difference or change in the CSF flow rate.

アガロース等電点電気泳動による神経変性疾患の髄液蛋白分画

The cerebrospinal fluid protein fractions were investigated by the agarose isoelectric focusing (agarose IEF) and the silver staining, for the purpose of elucidating protein abnormalities in the neural tissues in the degenerative neurological diseases, especially in motor neuron disease (MND), spinocerebellar degeneration (SCD) and Parkinson disease (PD).In this study, we focused six bands, mainly composed of transferrin, separated by agarose IEF and the silver staining. Some of these bands showed double fractions, even in the cases of the control group consisted of the patients without organic disease. In MND, all patients had double fractions of one or more of these bands, and it was interesting that in some of them double fractions were seen on the band with the isoelectric point of 5.6. Double fractions were also seen in many of SCD (69%) and PD (91%). Cases in that there was positive in more alkaline zone than transferrin in this method, were more in disease groups (especially in MND and SCD) than in the control group.This method combined agarose IEF with the silver staining seemed to be the excellent one as the investigating procedure for cerebrospinal fluid proteins.

Studies concerning the function of the complex subcommissural organ-liquor fibre: The binding ability of the liquor fibre to pyrocatechin derivatives and its functional aspects

(1) The subcommissural organ (SCO) and its secretion, condensed in the cerebrospinal fluid (CSF) to the Liquor fibre (LF, = Reissner's fibre), form a complex with still doubtful function. Recent results favour the concept of a detoxicating function of the SCO secretion in the CSF (binding hypothesis) 17,26,27,52. This study amplifies these results on the binding properties of LF to norepinephrine (NE) and epinephrine (E). The physiological significance of these properties is discussed. (2) After incubation of isolated bovine LF in solutions containing [ 3H]alanine or [ 3H]tyrosine and subsequent repeated washing, no radioactivity was detected by liquid scintillation counting in LF specimens. Hence, these substances cannot be firmly bound by bovine LF at the concentrations used. (3) When LF was incubated in solutions containing [ 14C]NE and/or [ 131I]-human serum albumin (HSA) in different concentration ratios and measured as above, no influence of HSA on the amount of amine bound to LF, nor of NE on the HSA amount was observed. The amount of HSA remaining bound to LF after washing was always very small, whereas that of NE exceeded its concentration in the surrounding medium. (4) Protein fractions of bovine CSF, incubated in solutions of [ 14C]NE, and separated by disc electrophoresis, contained no radioactivity. Hence we suggest that, in comparison with these proteins (and with HSA, see above), the binding properties described are also LF specific. (5) Catecholamines, capable of disturbing the activity of the nervous system, were found in the CSF in variable amounts ( e.g. see refs. 11, 13, 14, 44). Whether the binding properties of the Liquor fibre have some physiological significance is therefore discussed. The most favourable conditions for such an assumption seem to exist, for various reasons, in the central canal of the spinal cord.

Electrophoretic analysis of cerebrospinal fluid proteins in patients with central nervous system mass lesions.

✓ The protein constituents of the cerebrospinal fluid (CSF) from 234 patients with central nervous system mass lesions and nine patients with pseudotumor syndrome were analyzed by cellulose acetate electrophoresis. The alpha 2 and gamma globulin fractions were most frequently elevated, and more so with metastatic and malignant glial tumors than with benign tumors. Elevation of either fraction frequently occurred independent of the total protein content of the CSF and appears to indicate a primary parenchymal involvement of the brain. Gamma globulin elevations in patients with benign glial tumors were often associated with intracerebral cyst formation. Pituitary adenomas were not associated with elevation of the gamma globulin fraction whereas this was a common finding with other parasellar lesions. All CSF specimens associated with masses of infectious origin had markedly elevated gamma globulin levels irrespective of normal total protein levels. Two metastatic tumors from the gastrointestinal tract, a solitary plasmacytoma, and a case of increased intracranial pressure secondary to nalidixic acid therapy showed dramatic and specific increases in certain CSF protein fractions. The origin of the various protein fractions is discussed as well as the overall clinical applicability of CSF protein electrophoresis in patients with intracranial mass lesions.

The Protein Fractions in Cerebrospinal Fluid in the Various States of Multiple Sclerosis

(1) Agar gel electrophoresis was performed on the CSF of 139 patients with multiple sclerosis of both the intermittent and chronic type, and in the early and late stages of the disease. In most cases

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Analysis of Protein Fractions of Cerebrospinal Fluid by Means of Disc-electrophoresis (Polyacrylamide-Gel Electrophoresis)

Fractionation of normal cerebrospinal fluid proteins was carried out by Disc-electrophoresis and the obtained patterns were compared with those of normal sera.Protein fractions of cerebrospinal fluid were named provisionally as follows:Prealbumin (ρ), albumin (A), αL-fraction, α1'-and α2'-fraction, transferrin (Tr), β2'-fraction, βγ-fraction, and Sα2-fraction. The mean value (17 cases) of relative concentration of each protein fraction in normal cerebrospinal fluid was as follows; ρ: 4.0%, A: 73.7%, αL: 2.1%, α1'-α2': 3.8%, Tr: 5.8%, β2': 3.0%, βγ: 5.6%, Sα2: 2.0%.

Chronic Hypervitaminosis A in Holstein Male Calves

Summary Beginning at two and one-half months of age, daily vitamin A intakes from 62.5 through 16,000 μ g per pound of live weight were fed to 28 calves for 12 wk. At intakes (estimated by regression) above 3,981 μ g the ration consumed decreased and above 3,311 live weight gain decreased. Characteristic signs of hypervitaminosis A were observed in calves fed an 8,000 intake or above. Heart rate was elevated at intakes greater than 1,820 and cerebrospinal fluid (CSF) pressure decreased above 1,445. Vitamin A concentrations in plasma increased across all intakes and in liver up to an intake of 5,623, thereafter decreasing. Plasma ascorbic acid decreased across all intakes and tocopherol to 2,399, but thereafter increased slightly. The range of intakes above which increases or decreases were estimated to occur for serum protein fractions, inorganic phosphorus, and transaminase (GOT and GPT) and alkaline phosphatase activities and prothrombin time were from 1,000 through 4,365. At intakes exceeding 99, CSF protein fractions were affected. Liver, heart, and kidney weights per unit live weight increased above intakes of 1,995, 3,981, and 3,467. Metacarpal bones exhibited change in composition at intakes greater than 603.

Cerebrospinal fluid protein fractions in health and disease.

Samples of lumbar cerebrospinal fluid from 20 healthy subjects and 20 patients with a variety of neurological disorders have been subjected to cellulose acetate electrophoresis and quantitative analyses carried out on the seven separated protein fractions. Normal percentage and absolute concentration limits for each fraction have been determined and compared with previous reports. It is concluded that absolute values are more valuable than percentage values in the discrimination of disease from health, but that the mechanisms governing protein abnormalities are difficult to ascertain from spinal fluid electrophoresis alone.

Changes of the cerebrospinal fluid protein picture in polyradiculoneuritis

Foi estudado por eletroforese em papel o proteinograma do LCR de 7 pacientes com polirradiculoneurite; em 5 foi estudado também o proteinograma do sôro pelo mesmo método. As alterações verificadas permitem caracterizar o proteinograma do LCR nesta afecção como sendo de tipo misto, acompanhado de aumento de a-globulinas, segundo a classificação de Bauer. Foi mostrado que as modificações observadas no teor das diversas frações protêicas no LCR podem ser secundárias a alterações verificadas no sôro e que podem decorrer de distúrbios do mecanismo de ultrafiltração das proteínas do sangue para o LCR, por modificações da barreira hêmato-liquó-rica. O comportamento particular da a1globulina é salientado; seu teor estava aumentado no LCR em 6 casos e, em relação ao teor desta globulina no proteinograma do sôro, sua participação no do LCR chegou a ser quase 5 vêzes maior. Para explicar esta observação, além da hipótese anterior, é lembrada a de poderem ocorrer no LCR, nos casos de polirradiculoneurite, modificações estruturais desta globulina e/ou das relações entre ela e as substâncias de que é carreadora, modificando sua reabsorção no sentido de se produzir uma estase relativa.

Eletroforese das proteínas do líqüido cefalorraquidiano na cisticercose do sistema nervoso central

In order to analyse the paper strip electrophoresis contribution to the knowledge of cerebrospinal fluid (CSF) proteins in cysticercosis of the central nervous system (CNS) 40 patients were studied (identification data in table 1); the clinical forms and diagnostic data of 30 of them, who had CNS cysticercosis (cases 1 to 30) are summarized in the table 2; cysticercosis of the CNS might play a role in the pathologic condition presented by the remaining 10 patients. CSF protein fractions were examined in all the cases (results in tables 4 and 7) and those of blood sera in cases from 1 to 15 (results in table 5) by paper strip electrophoresis under the specifications previously reported54. A second examination of CSF protein fractions was made some time later in 4 cases (results in table 6). CSF samples were analysed also in respect to cytology, total protein content, Pandy and colloidal benzoin reactions and complement fixation tests for syphillis and cysticercosis (results in table 3). The values obtained for CSF protein fractions were compared to norma) values found in 30 control subjects; these normal values were detailed in a prior publication55 and are summarized in table 8. This comparison shows that CNS cysticercosis produces changes in the electrophoretic prophiJe of CSF proteins (table 9); an increased y-globulin fraction was the main change observed, commonly associated to a low j8-globulin relative concentration. An inversion of a1/a2 quocient was found in 4 cases. The contributions to the study of CNS cysticercosis resulting from the electrophoretic analysis of CSF proteins are discussed in three groups: 1 - Informations obtained by paper electrophoresis of CSF proteins are independent from those resulting from the other laboratory aspects studied. Thus, if the protein fractions of blood sera are considered, although the changes found have mean values similar to those found to the CSF protein fractions (table 10), it was observed that there are individual differences in the protein fraction relative concentrations in blood and CSF (table 11), no correlation being found between them. Considering CSF itself, changes in its protein fractions induced by CNS cysticercosis are similar in ventricular and subarachnoid samples (lumbal and cisternal) (table 13); their intensity is related to the intensity of the whole of CSF changes (table 12). Informations obtained by electrophoretic analysis of CSF proteins, however, differ from those resulting from the other examinations conducted in the CSF sample as was shown in the analysis of cytology (tables 14 and 15), total protein content (table 16), colloidal benzoin reaction and positivity of complement fixation test for cysticercosis. 2 - The clinical value of data achieved by CSF proteins electrophoresis is discussed. Results may have an important role in diagnosis, in the knowledge of its clinical forms (table 17) and in the control of evolution if pathogenetic mechanisms involved in disease are considered. Cases 31 to 40 illustrate the clinical aspects of contributions given by CSF electrophoresis. 3 - The CSF protein fractions changes found in CNS cysticercosis justify their classification among those changes commonly observed in subchronic and chronic inflammatory diseases of the CNS and/or its leptomeningeal coverings. It is assumed in the literature that there occurs a local production of globulins, specially the y-globulin fraction, in such pathologic conditions. Concerning CNS cysticercosis, if CSF total protein content and electrophoretic data on its fractions are considered together (table IS and 19; graph. 1), it is possible to evidence that at least two mechanisms participate in the origin of protein changes (tables 20 and 21; graph. 2) : blood-CSF barrier disturbances are able to explain data concerning albumin, a and globulin changes, which are similar; the local production of y-globulin is the hypothesis most reliable to explain the peculiar changes of this fraction. This hypothesis agrees with literature data concerning other chronie inflammatory diseases of the CNS. The probable role of y-globulin in carrying specific antibodies is pointed out through correlative exploration of its concentration in the CSF sample and the corresponding positivity of complement fixation test for cysticercosis.

Abnormal protein fractions of cerebrospinal fluid demonstrated by starch gel electrophoresis.

Abnormal protein fractions of cerebrospinal fluid demonstrated by starch gel electrophoresis.

Protein studies in normal newborn infants; paper electrophoresis of cerebrospinal fluid and serum.

Serum protein fractions have been studied intensively in the normal newborn infant. 1-9 However, there is only sparse information on the cerebrospinal fluid protein fractions in this age group. 10-18 The concentration of the low protein content in small amounts of cerebrospinal fluid has presented a major problem. Earlier studies primarily used precipitation methods which could not separate the individual fractions. The use of dialysis methods for concentrating proteins, combined with paper electrophoresis, now affords adequate means for separation of the various components of cerebrospinal fluid proteins. Paper electrophoretic studies of cerebrospinal fluid have been performed for the most part only in adults and older children, 19-22 with few reports in normal newborn infants. 17,18 The purpose of this study was to determine the distribution and relationship of proteins in the cerebrospinal fluid and serum of normal newborn infants utilizing chemical and paper electrophoretic methods. Method Thirty-five normal newborn infants

Electrophoresis of cerebrospinal proteins. I. General findings on paper electrophoresis

Os caracteres gerais do método eletroforético são apresentados de modo sumário, especialmente aquêles referentes à eletroforese em papel, no sentido de familiarizar o neurologista quanto aos seus aspectos principais e sua utilização no estudo das frações proteicas do liqüido cefalorraquidiano cujos aspectos particulares são discutidos.