Prognostic relevance of the C-X-C motif chemokine ligand 13 and interleukin-8 in predicting the transition from clinically isolated syndrome to multiple sclerosis.

The initial phase of multiple sclerosis (MS), often known as clinically isolated syndrome (CIS), is a critical period for identifying individuals at high risk of progressing to full-blown MS and initiating timely treatment. In this study, we aimed to evaluate the prognostic value of C-X-C motif chemokine ligand 13 (CXCL13) and interleukin-8 (IL-8) as potential markers for CIS patients' conversion to MS. Our study encompassed patients with CIS, those with relapsing-remitting MS (RRMS), and control subjects, with sample analysis conducted on both cerebrospinal fluid (CSF) and serum. Patients were categorized into four groups: CIS-CIS (no MS development within 2 years), CIS-RRMS (conversion to RRMS within 2 years), RRMS (already diagnosed), and a control group (CG) with noninflammatory central nervous system disorders. Results showed significantly elevated levels of CXCL13 in CSF across all patient groups compared with the CG (p < 0.0001, Kruskal-Wallis test). Although CXCL13 concentrations were slightly higher in the CIS-RRMS group, statistical significance was not reached. Similarly, significantly higher levels of IL-8 were detected in CSF samples from all patient groups compared with the CG (p < 0.0001, Kruskal-Wallis test). Receiver operating characteristic analysis in the CIS-RRMS group identified both CXCL13 (area under receiver operating characteristic curve = .959) and IL-8 (area under receiver operating characteristic curve = .939) in CSF as significant predictors of CIS to RRMS conversion. In conclusion, our study suggests a trend towards elevated CSF IL-8 and CSF CXCL13 levels in CIS patients progressing to RRMS. These findings emphasize the importance of identifying prognostic markers to guide appropriate treatment strategies for individuals in the early stages of MS.

Cytological examination of cerebrospinal fluid.

Cytological examination of cerebrospinal fluid (CSF) has some specifics. In the pre-analytical phase, due to collection into a tube without fixation and hypooncotic nature with rapid cell degradation, processing is required within 2-3 hours. In the subsequent analytical phase it is necessary to take into account the quantitative limit of the sample due to regular parallel cytological processing and non-morphological methods. A regular entry step of cytological examination is the quantification of cellularity. Even oligocellular samples may exhibit severe cytological findings. In the category of inflammatory processes, the cytological diagnosis of purulent meningitis within a few hours has a fundamental influence on prognosis of the disease. Non-purulent inflammations with a broad differential diagnosis are specified not only by cytological images, but also by parallel non-morphological examinations aimed at detecting an infectious agent or autoimmune nature of inflammation. Neoplastic processes in the intermeningeal space generally require immunocytochemical verification. The minimum amount of CSF for immunocytochemical examinations is 3-5 ml. The requirement for these examinations resulting from the baseline assessment may indicate the need for another sample. If the appropriate procedures and interpretations are followed in the context of parallel non-morphological examinations, the cytological examination of MM can contribute significantly to the diagnosis of pathological conditions in CSF spaces.

Multicentre comparison of a diagnostic assay: aquaporin-4 antibodies in neuromyelitis optica.

OBJECTIVE: Antibodies to cell surface central nervous system proteins help to diagnose conditions which often respond to immunotherapies. The assessment of antibody assays needs to reflect their clinical utility. We report the results of a multicentre study of aquaporin (AQP) 4 antibody (AQP4-Ab) assays in neuromyelitis optica spectrum disorders (NMOSD). METHODS: Coded samples from patients with neuromyelitis optica (NMO) or NMOSD (101) and controls (92) were tested at 15 European diagnostic centres using 21 assays including live (n=3) or fixed cell-based assays (n=10), flow cytometry (n=4), immunohistochemistry (n=3) and ELISA (n=1). RESULTS: Results of tests on 92 controls identified 12assays as highly specific (0-1 false-positive results). 32 samples from 50 (64%) NMO sera and 34 from 51 (67%) NMOSD sera were positive on at least two of the 12 highly specific assays, leaving 35 patients with seronegative NMO/spectrum disorder (SD). On the basis of a combination of clinical phenotype and the highly specific assays, 66 AQP4-Ab seropositive samples were used to establish the sensitivities (51.5-100%) of all 21 assays. The specificities (85.8-100%) were based on 92 control samples and 35 seronegative NMO/SD patient samples. CONCLUSIONS: The cell-based assays were most sensitive and specific overall, but immunohistochemistry or flow cytometry could be equally accurate in specialist centres. Since patients with AQP4-Ab negative NMO/SD require different management, the use of both appropriate control samples and defined seronegative NMOSD samples is essential to evaluate these assays in a clinically meaningful way. The process described here can be applied to the evaluation of other antibody assays in the newly evolving field of autoimmune neurology.

Biochemical manifestations of the nervous tissue degradation after the blood-brain barrier opening or water intoxication in rats.

OBJECTIVES: The aim of the study was to determine changes of biomarkers of nervous tissue degradation in experimental model of osmotic blood-brain barrier opening or water intoxication and to find whether they correspond to changes in well defined clinical entities. METHODS: In the cerebro-spinal fluid taken via the suboccipital puncture, myelin basic protein (MBP ng/ml), neuron-specific enolase (NSE ng/ml) and TAU-protein (Tau pg/ml) were determined by ELISA in 19 controls and 29 experimental rats several hours or one week after the experimental intervention. RESULTS: Significant difference between the control and experimental groups was revealed only for the concentration of myelin basic protein. After the BBB opening, its level dramatically increased within hours and dropped back to control values within one week. Water intoxication induced only dilutional hypoproteinorachia. No significant changes were found in NSE and levels of TAU-protein were not detectable. CONCLUSION: 1. Increased permeability of cytoplasmic membranes induced by water intoxication does not alter any of monitored CSF biomarkers. 2. Osmotic opening of the BBB in vivo experiment without the presence of other pathological conditions leads to a damage of myelin, without impairment of neurons or their axons.

IL-2, IL-6 and chitinase 3-like 2 might predict early relapse activity in multiple sclerosis.

BACKGROUND: The possibility to better predict the severity of the disease in a patient newly diagnosed with multiple sclerosis would allow the treatment strategy to be personalized and lead to better clinical outcomes. Prognostic biomarkers are highly needed. OBJECTIVE: To assess the prognostic value of intrathecal IgM synthesis, cerebrospinal fluid and serum IL-2, IL-6, IL-10, chitinase 3-like 2 and neurofilament heavy chains obtained early after the onset of the disease. METHODS: 58 patients after the first manifestation of multiple sclerosis were included. After the initial diagnostic assessment including serum and cerebrospinal fluid biomarkers, all patients initiated therapy with either glatiramer acetate, teriflunomide, or interferon beta. To assess the evolution of the disease, we followed the patients clinically and with MRI for two years. RESULTS: The IL-2:IL-6 ratio (both in cerebrospinal fluid) <0.48 (p = 0.0028), IL-2 in cerebrospinal fluid ≥1.23pg/ml (p = 0.026), and chitinase 3-like 2 in cerebrospinal fluid ≥7900pg/ml (p = 0.033), as well as baseline EDSS ≥1.5 (p = 0.0481) and age <22 (p = 0.0312), proved to be independent markers associated with shorter relapse free intervals. CONCLUSION: The IL-2:IL-6 ratio, IL-2, and chitinase 3-like 2 (all in cerebrospinal fluid) might be of value as prognostic biomarkers in early phases of multiple sclerosis.

Assisting the neurologist in diagnosis of CNS malignancies - Current Possibilities and Limits of Cerebrospinal Fluid Cytology and Immunocytochemistry.

OBJECTIVES: In tumorous impairment of CNS, cytological identification of the neoplastic cells in CSF frequently requires the use of ancillary techniques. Our methods are focused on identifying algorithms that increase the probability of identifying CSF malignant cells. MATERIALS AND METHODS: A total of 1.272 CSF samples from patients with tumorous infiltration of CNS of nonhematologic origin along with 721 samples from patients with hematologic malignancies were analyzed in a complex setting including cytological and immunocytochemical investigations. RESULTS AND DISCUSSION: In CSF diagnostics we are aware of the limited amount of sample combined frequently with neoplastic oligocytosis. Provided atypical, potentially malignant cells in CSF are found, further investigation(s) should maximize the probability of their identification-an appropriate cytological staining and immunocytochemical panel is to be applied. (i) In cases of known recent malignancy: immunoprofile of the recent neoplasm has been considered in immunocytochemical panel. (ii) In patients with a history of malignancy: The propensity to develop a new different malignancy must be taken into account. (iii) Atypical cells found in the CSF of a patient with a negative history of malignancy: Considering the most frequent clinically silent malignancies, stepwise immunocytochemistry is employed. Three milliliter of initial CSF sample represents the absolute minimum to start with. CONCLUSIONS: The steps of the laboratory activity targeted on malignancy in the CSF detection can be expected as follows: (i) The sample will be divided for both nonmorphology and cytopathology investigations. (ii) Basic stainings will triage the samples into those with no suspicion of malignancy and the remaining ones. (iii) Special stainings and stepwise immunocytochemistry will be performed in parallel with the nonmorphology investigations.

Biomarkers of Brain Damage: S100B and NSE Concentrations in Cerebrospinal Fluid--A Normative Study.

NSE and S100B belong among the so-called structural proteins of the central nervous system (CNS). Lately, this group of structural proteins has been profusely used as specific biomarkers of CNS tissue damage. So far, the majority of the research papers have focused predominantly on the concentrations of these proteins in blood in relation to CNS damage of various origins. Considering the close anatomic and functional relationship between the brain or spinal cord and cerebrospinal fluid (CSF), in case of a CNS injury, a rapid and pronounced increase of the concentrations of structural proteins specifically in CSF takes place. This study inquires into the physiological concentrations of NSE and S100B proteins in CSF, carried out on a sufficiently large group of 601 patients. The detected values can be used for determination of a normal reference range in CSF in a clinical laboratory diagnostics.

CSF and serum orosomucoid (alpha-1-acid glycoprotein) in patients with multiple sclerosis: a comparison among particular subgroups of MS patients.

INTRODUCTION: To compare cerebrospinal fluid (CSF) and serum orosomucoid (alpha-1-acid glycoprotein-AAG) concentrations in various subgroups of patients with multiple sclerosis (MS). MATERIALS AND METHODS: CSF and serum AAG concentrations, AAG quotient (i.e., CSF AAG/serum AAGx10(3)) and index were determined in a group of 59 patients with clinically definite or probable MS. Patients were subdivided according to the disease form, disease severity according to an expanded disability status scale (EDSS), its treatment, disease duration and sex. RESULTS: CSF AAG was increased in 52.5% of the patients and AAG quotient even in 64.4%. An increase in the CSF AAG concentration, as well as in AAG quotient and index, appear only after several years of disease duration, while no significant correlation with age has been found. This suggests that CSF AAG changes in MS represent a secondary, unspecific phenomenon and that this protein is not relevant for the aethiopathogenesis of the disease. Nevertheless, the finding of subnormal CSF AAG levels in some MS patients in remission (never observed in those in the attack) implies the possibility that CSF AAG may be used as a "state marker" in MS. Serum AAG levels were significantly lower in secondary progressive form and in severely disabled patients. This observation suggest that serum AAG values determination might have some prognostic significance. Further studies are, however, needed. Serum AAG should be investigated in parallel with other CSF and serum protein fractions in order to establish a pannel of examinations enabling multiple statistical analyses. This approach may lead to the finding of a "complex state marker" enabling thus to evaluate more precisely disease course in individual patients and to accept appropriate therapeutic measures.

Coefficient of energy balance: effective tool for early differential diagnosis of CNS diseases.

Urgent examination of cerebrospinal fluid (CSF) provides immediate important information about the character of central nervous system (CNS) impairment. Although this examination includes energy parameters such as glucose and lactate concentrations, it does not commonly use Coefficient of Energy Balance (CEB). In this study, we focused on CEB because it enables more exact assessment of actual energy state in the CSF compartment than glucose and lactate alone. CEB informs about the actual functioning condition of present cells, and it does not require any other analysis or costs. Using Kruskal-Wallis ANOVA, we examined a large CSF sample (n = 8183) and we compared CEB values among groups with different cytological syndromes. We found a statistically significant difference of CEB between the group with granulocyte pleocytosis and the control group. These results indicate a high degree of anaerobic metabolism caused by the oxidative burst of neutrophils. Similarly, we found a statistically significant difference of CEB between the control group and groups with tumorous oligocytosis plus pleocytosis and monocyte pleocytosis. This difference can be attributed to the oxidative burst of macrophages. Our findings suggest that CEB combined with CSF cytology has a great importance for diagnosis, differential diagnosis, and early therapy of CNS diseases.