Signs of myelin impairment in cerebrospinal fluid after osmotic opening of the blood-brain barrier in rats.

A number of clinical neurological pathologies are associated with increased permeability of the blood brain barrier (BBB). Induced changes of the homeostatic mechanisms in the brain microenvironment lead among others to cellular changes in the CNS. The question was whether some of these changes can be induced by osmotic opening of BBB in an in vivo experiment and whether they can be detected in cerebrospinal fluid (CSF). CSF was taken via the suboccipital puncture from 10 healthy rats and six rats after the osmotic opening of the BBB. In all 16 animals, concentration of myelin basic protein (MBP ng/ml), Neuron-specific enolase (NSE ng/ml) and Tau-protein (Tau pg/ml) were determined in CSF by ELISA. Values in both groups were statistically evaluated. Significant difference between the control and experimental group was revealed only for the concentration of myelin basic protein (p<0.01). The presented results indicate that osmotic opening of the BBB in vivo experiment without the presence of other pathological conditions of the brain leads to a damage of myelin, without impairment of neurons or their axons.

Immunophenotypic analysis of cerebrospinal fluid cell populations with the Cell-Dyn Sapphire haematology analyser: method feasibility and preliminary observations.

Cerebrospinal fluid (CSF) samples (n=50) from patients with neurological disease (bacterial infection, viral infection, neuroborreliosis and multiple sclerosis) were analysed to characterize cell populations by fluorescent immunocytometry with the CD-Sapphire haematology analyser. Reagent combinations applied to all CSF samples comprised CD3/CD19/HLA-DR and CD4/CD8, with some being further analysed using CD3/CD4, CD3/CD16 and CD3/CD25 protocols. Of the 50 samples, 11 were excluded because of high proportions of nonviable cells (n=2) or insufficient cell numbers (n=9). Apart from bacterial infection with granulocytosis, all diagnostic groups showed high proportions (51.4-77.0%) of CD3+ T cells. There was a modest association between T-cell and B-cell counts, but absolute B-cell numbers exceeded 5 cells/microl in only 7/39 cases (neuroborreliosis, n=6; bacterial meningitis, n=1). CD3/Ia antigen (activation) co-expression was low and only exceeded 5% in 7/39 samples with no diagnostic correlation. Primary CD4+ and CD8+ T-cell subsets showed similar quantitative trends and CD4/CD8 co-analysis revealed the presence in all diagnostic groups (neuroborreliosis and multiple sclerosis in particular) of a CD4+CD8int fraction that was predominantly CD3+ and CD16- and had a morphological profile consistent with small lymphoid cells. Supplementary CD-Sapphire cellular immunological analysis of most CSF samples is feasible using the procedure detailed in this communication.

Eosinophilic meningitis - an immunophenotyping recording of a very rare clinical entity - brief Report.

A very rare clinical entity, so-called eosinophilic meningitis, classified by prevalence of eosinophils in cerebrospinal fluid (CSF), with the presence of pleiocytosis, has been recorded in our laboratory four times only in the last 24 years. A low glucose level, elevation of total protein and lactic acid in CSF were detected in all the clinical cases. The last two cases were made possible by using flow cytometry method; surprisingly, the presence was found in mature T-cells in CSF, predominantly helpers (CD3+, CD4+) and, practically, none is B-cells (CD19+), plasma cells (CD138+) and NK-cells.

Cerebrospinal fluid and serum prealbumin (transthyretin) in patients with multiple sclerosis (MS): comparison of particular subgroups of MS patients.

The levels of prealbumin (PAB, transthyretin) were determined and evaluated in the cerebrospinal fluid (CSF) and serum in various subgroups of the multiple sclerosis (MS) patients. In severely disabled patients, serum PAB was elevated more frequently. CSF and serum PAB concentrations were higher in treated than in nontreated patients; the values above the upper reference limits were also more frequently found in treated patients. PAB index showed a tendency to decrease during the course of the disease. The routine determination of PAB in CSF and serum is, therefore, recommended to be recognized in MS patients as a substantial clinical value and, thus, be comprised, including also immunoglobulins and other parameters, into the spectrum of characteristics in Protein Panel.

Cryptococcal meningitis--a follow-up study of a serious clinical entity: quick cytological and microbiological diagnostics using a special staining procedure in cerebrospinal fluid specimens.

A routine diagnostic procedure of cryptococcal meningitis using Alcian Blue and Nuclear Fast Red staining is described in a group of 16 patients. Cerebrospinal fluid findings, including clinical cytology, routine biochemistry and protein fractions, are presented.

Complete proteinogram of cerebrospinal fluid and its contribution to the diagnostics of inflammatory and autoimmune diseases of the central nervous system.

Complete evaluation of cerebrospinal fluid proteinogram represents a routine request of the clinician in the analysis of CSF in the Czech Republic. It comprises the measurement of concentrations of acute phase proteins (CRP, orosomucoid, haptoglobin, transferrin, prealbumin), immunoglobulins (IgG, IgA, IgM), compressive markers (albumin, fibrinogen), markers of CNS tissue destruction (apolipoproteins A-I, A-II, Apo B), complement components (C3, C4), alpha-1-microglobulin, beta-2-microglobulin, and proteinase inhibitors (alpha-1-antitrypsin, antithrombin III). Therefore, 19 CSF proteins of precisely verified clinical relevance are routine parameters for the assessment of the functional state of the blood-CSF barrier, presence of the intrathecal synthesis of immunoglobulins, inflammatory changes and verification of CNS tissue destruction. Evidence of these clinically relevant and independent parameters enabled the detection of the presence of autoimmune and neuroinfective diseases of CNS, even in clinical cases where the basic CSF parameters do not express relevant changes, or they are of a bordering or non-specific character. Clinically typical and the most significant abnormalities in the CSF proteinogram represent themselves a new access to a contemporary CSF analysis. Despite the fact that assessment of CSF proteins and their analysis is quite a difficult field in laboratory medicine, it is now routinely requested and routinely performed in the Czech Republic.

Levels of apolipoprotein A-II in cerebrospinal fluid in patients with neuroborreliosis are associated with lipophagocytosis.

Levels of most of the examined proteins in cerebrospinal fluid (CSF) of 107 patients with neuroborreliosis were associated with cytological findings, the status of hematoencephalic barrier as evaluated by Qalb (cerebrospinal fluid to serum quotient) and the intrathecal synthesis of immunoglobulins. Cytological findings consisted of normal cytology, or both oligocytosis and pleocytosis of monocytes or lymphocytes. The lipophagic elements were present in 20% of samples. Concentrations of apolipoproteins A-I and A-II in the CSF were correlated with the concentration of albumin without regard to the CSF cytology. The levels of apolipoprotein B were increased only in samples with lymphocytic pleocytosis and Qalb > 7.4. The presence of lipophages in the CSF was significantly associated with the CSF concentration of apolipoprotein A-II.

Cerebrospinal fluid cytologic findings in multiple sclerosis. A comparison between patient subgroups.

OBJECTIVE: To compare cytologic findings in cerebrospinal fluid (CSF) in various subgroups of multiple sclerosis (MS) patients. STUDY DESIGN: CSF from 77 patients with clinically definitive or probable MS was examined by means of qualitative cytology. After the cell count was determined in a Fuchs-Rosenthal chamber, slides were prepared by the cytosedimentation method and stained with May-Grünwald-Giemsa stain and oil red O and, whenever possible, with Papanicolaou stain and toluidine blue. In addition to the differential cell count, the lymphocyte/monocyte ratio, percentage of activated forms in the lymphocytic and monocytic series, presence and percentage of lymphoplasmacytes and mature plasma cells, presence of lipophages, lymphophages and presence of mitotic figures were evaluated. RESULTS: The following statistically significant differences were found between the various MS subgroups: (1) higher prevalence of mitotic figures in the primary progressive MS subgroup; (2) higher prevalence of foam cells and lymphophages and lower prevalence of CSF pleocytosis in more severely disabled patients; (3) lower cell count, lower prevalence of CSF pleocytosis, lower lymphocyte/monocyte ratio and lower prevalence of lymphoplasmacytes in treated patients; and (4) higher prevalence of mature plasma cells and lipophages in MS patients with disease of longer duration. CONCLUSION: The differences observed in the various MS subgroups may reflect certain aspects of MS pathogenesis. Qualitative CSF cytology may therefore be useful for both clinicians and neuroimmunologists. Qualitative cytology of CSF is an important diagnostic method that should never be omitted from an examination of CSF from patients with MS.

Transferrin in patients with multiple sclerosis: a comparison among various subgroups of multiple sclerosis patients.

OBJECTIVES: To compare cerebrospinal fluid (CSF) and serum transferrin (Tf) concentrations, transferrin quotient and index in various subgroups of MS patients. MATERIAL AND METHODS: CSF and serum transferrin concentrations, transferrin quotient QTf (i.e. CSF transferrin/serum transferrin x 10(3)) and index (QTf/Qalbumin) were determined in a group of 51 patients with clinically definite or probable multiple sclerosis (MS). Patients were subdivided according to the disease form (relapsing-remitting = RR, secondary progressive = SP, primary progressive = PP; patients with RR form were further subdivided into those in the attack and those in remission), disease severity (EDSS 0-5.5, EDSS 6.0-10.0), its treatment (non-treated - including patients treated with vitamins and/ or vasodilators only, treated - i.e. glucocorticoids and/or immunosuppressants and/or (exceptionally) beta-interferon), disease duration (0-2 years, >2-10 years, > 10 years) and sex. Correlation of transferrin values with age was also performed. RESULTS: Serum transferrin was somewhat lower and significantly more frequently subnormal in PP patients in comparison with the SP form and the RR form in remission. Transferrin index was significantly higher in the PP form than in the RR as well as the SP form. Transferrin quotient was significantly more frequently subnormal in patients in remission compared to those in the attack of the RR disease. CSF transferrin as well as transferrin quotient were more frequently subnormal in patients with short disease duration (0-2 years) than in patients with longer disease duration; these parameters, however, correlated also significantly with age. CSF transferrin and transferrin quotient were higher in male than in female patients. CONCLUSION: The authors conclude that evaluation of transferrin in MS patients - along with albumin - may help to differentiate among various MS subgroups, since there are significant differences among RR, SP and PP forms. For this purpose, however, other CSF protein fractions should be evaluated in parallel in order to obtain more complex information and to establish a panel of examinations enabling multiple statistical analyses. Transferrin evaluation in MS may also be of significant theoretical interest, since transferrin is known to be involved in the regulation of iron metabolism and it may have a protective role against the oxidative stress. Moreover, transferrin is a growth factor important for proliferation of activated T lymphocytes. By means of the use of transferrin quotient and especially transferrin index, it may be possible to estimate the proportion of intra-CNS-synthesized transferrin and/or rate of specific transferrin transport across the blood-CSF barrier. Further studies are, however, needed for such an evaluation.