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Determination of cefepime in plasma and cerebrospinal fluid by micellar electrokinetic chromatography with direct sample injection.

A simple micellar capillary electrokinetic chromatography (MEKC) with UV detection is described for analysis of cefepime in plasma and cerebrospinal fluid by direct injection without any sample pretreatment. The separation of cefepime from biological matrix was performed at 25 degrees C using a background electrolyte consisting of tris(hydroxymethyl)aminomethane (Tris) buffer with sodium dodecyl sulfate (SDS) as the electrolyte solution. Under optimal MEKC condition, good separation with high efficiency and short analyses time is achieved. Several parameters affecting the separation of the drug were studied, including the pH and concentrations of the Tris buffer and SDS. Using cefazolin as an internal standard, the linear ranges of the method for the determination of cefepime in plasma and cerebrospinal fluid were 1-50 and 1-20 microg/mL, respectively; the detection limits of plasma (signal-to-noise ratio = 3; injection, 5 kV, 5 s) and cerebrospinal fluid (signal-to-noise ratio = 3; injection, 0.5 psi, 3 s) were 0.2 microg/mL and 0.3 microg/mL, respectively. Application of the proposed method for determination of cefepime in plasma and cerebrospinal fluid collected after intravenous administration of 2 g cefepime in patients with meningitis was demonstrated.

Cefepime↗

[Characteristic cerebrospinal fluid findings and clinical aspects of lymphocytic meningoradiculitis].

Clinical data and Cerebrospinal fluid (CSF) findings of 20 patients suffering from a meningoradiculitis were analyzed. Most patients had a tick bite or an erythema migrans before neurological symptoms occurred. All suffered from severe radicular pain. Predominant neurological symptoms were peripheral palsies of the facial nerve followed by motor paresis of the trunk and the extremities of mostly asymmetric distribution. Predominant CSF alterations were a long lasting inflammatory cellular reaction and an impairment of the blood-CSF barrier. In contrast to the CSF of 17 patients suffering from a polyradiculitis Guillain Barré with a similar total protein content, an intrathecal immunoglobulin IgG synthesis could be confirmed for the majority of the patients. CSF finding would be compatible to a viral as well as a borrelia etiology.

Bites and Stings↗

Radionuclide estimation of cerebrospinal fluid shunt flow. Evidence supporting an alternative theoretical model.

Flow of cerebrospinal fluid through a surgically implanted valve may be estimated by analyzing the disappearance curve resulting from the injection of a radiotracer into the valve. The standard method for estimating flow assumes an exponential disappearance of the tracer from the valve. This method models the valve as a single well-mixed compartment. Experimental evidence, showing that estimates of flow were dependent upon the site of injection, is at variance with this assumption. An alternative method of analyzing the disappearance curves, based on the area to height ratio (A/H) of the curves, was found to be more consistent with the experimental evidence and resulted in greater precision than the exponential method. It was concluded that optimal results are obtained using the A/H method with a fixed injection technique.

Cerebral Ventriculography↗

Detection of Epstein-Barr virus DNA in cerebrospinal fluid from immunocompetent individuals with brain disorders.

Fifty four cerebrospinal fluid samples obtained from as many immunocomponent patients with disorders of the central nervous system were investigated for the presence of herpesvirus DNA by nested polymerase chain reaction in order to determine an etiological diagnosis. Four of these samples proved positive for the presence of Epstein-Barr virus DNA (7.4%). The result of this diagnostic study is reported to draw insiders' attention to the possible presence of EBV in cerebrospinal fluid from patients with central nervous system diseases.

Adult↗

Cerebrospinal fluid cells and proteins in patients occupationally exposed to organic solvents.

Cerebrospinal fluid (CSF) cells and proteins were determined for 33 patients exposed to industrial organic solvents. A lymphoid reaction, i.e., a pathologically elevated number or percentage of enlarged lymphoid cells was observed in one-third of the patients, more often in patients with chronic intoxication (40%) than in those currently exposed to organic solvents (32%). An almost significant decrease of small lymphocytes in the CSF was observed among patients who had a past history of chronic solvent intoxication but no recent exposure. No cytological evidence of tissue destruction was found. Signs of slight blood--CSF barrier damage occurred in 5 (23%) of the currently exposed patients, but intrathecal IgG synthesis was not observed. Increased cellular activity in the CSF was also accentuated in principal component analysis. The results suggest slight nonspecific immunoactivation in the central nervous system of subjects exposed to organic solvents.

Adult↗

Intracranial hypertension after treatment of spontaneous cerebrospinal fluid leaks.

Four patients, aged 10 to 44 years, with spontaneous cerebrospinal fluid (CSF) leaks and intracranial hypotension developed intracranial hypertension after treatment of their CSF leaks. The leak was at the spinal level in all patients (thoracic level, 2; lumbar level, 1; and undetermined, 1). One patient responded to an epidural blood patch. Three patients responded to surgery, of whom 2 had not responded to prior epidural blood patches. Treatment resulted in complete resolution of symptoms, including orthostatic headaches and disappearance of magnetic resonance imaging abnormalities. However, all patients later developed steady headaches different from their previous headaches. None had recurrence of magnetic resonance imaging abnormalities or any evidence of occlusion of cerebral venous sinuses. All had increased CSF opening pressures. One had bilateral papilledema, and another had no venous pulsations on examination of fundi. Follow-up was possible in 2 patients. One responded well to treatment with acetazolamide, and the other improved gradually and was asymptomatic within several months.

Adolescent↗

Cerebrospinal fluid changes after iopamidol and metrizamide myelography in clinically normal dogs.

Cerebrospinal fluid samples from 2 groups of clinically normal dogs were compared after iopamidol (n = 9) and metrizamide (n = 8) myelography. Iopamidol (200 mg of I/ml) and metrizamide (170 mg of I/ml) were administered by cerebellomedullary injection at dosage of 0.45 ml/kg of body weight. In dogs of both groups, postmyelographic CSF changes included high specific gravity, Pandy score, protein concentration, and WBC count. The high specific gravity and Pandy score were false-positive effects attributed to nonionic contrast media. Although postmyelographic protein concentration and total WBC count were greater in CSF samples from dogs given metrizamide than in those given iopamidol, differences were not statistically significant. The differential WBC counts were consistent with mild, acute leptomeningitis; these findings were supported by results of histologic examination. Iopamidol and metrizamide should be considered low-grade leptomeningeal irritants in dogs.

Animals↗

[Immunoglobulins in the cerebrospinal fluid of the newborn infant].

Immunoglobulins IgG, IgA and IgM were studied in serum and cerebrospinal fluid (CSF) samples from 36 newborn infants in the second day of life. According to parameters previously established in the same laboratory for normal newborn infants, serum proteins were normal in all samples and CSF composition was normal as to blood pigments, cells, total proteins and protein fractions in all samples. Immunoglobulins concentrations were determined by two methods: radial immunodiffusion (RID) and nephelometry. Concentrations found (mean +/- standard deviation) were as follows (mg/dl). Serum: through the RID method IgG 1346 +/- 299.1, IgM 10.4 +/- 2.26, IgA 1.1 +/- 1.06; through nephelometry IgG 1372 +/- 319.7, IgM 10.6 +/- 4.10, IgA 1.5 +/- 0.31. Cerebrospinal fluid: IgA and IgM were not detected by the methods employed; IgG was detected by both methods, and values were 11.1 +/- 2.52 by RID and 9.6 +/- 7.04 by nephelometry. IgG relations and IgG index were also evaluated; values of the IgG index (0.5 +/- 0.14) are emphasized. Considerations are made in order to point out data obtained as to characterize normal values for immunoglobulins in the cerebrospinal fluid of normal newborn infants.

Blood Protein Electrophoresis↗

In vitro cell-mediated immunity of cerebrospinal-fluid lymphocytes to myelin basic protein in primary demyelinating diseases.

In an attempt to characterize the immunologic reactivity of cerebrospinal-fluid lymphocytes in demyelinating diseases, we compared the myelin-basic-protein-induced in vitro responses of these cells to peripheral blood lymphocytes from the same subjects with a variety of neurologic diseases. Peripheral blood lymphocytes from patients with acute disseminated encephalomyelitis and progressive multiple sclerosis had increased reactivity as compared to those of normal volunteers (P less than 0.01 and P less than 0.05, respectively). Cerebrospinal-fluid lymphocytes from patients with acute disseminated encephalomyelitis and acute and progressive (but not stable) multiple sclerosis were more reactive than cells from subjects with other neurologic diseases (P less than 0.005, P less than 0.02 and P less than 0.05, respectively). Cerebrospinal-fluid lymphocytes manifested a greater reactivity than peripheral blood lymphocytes in acute and progressive multiple sclerosis but not in acute disseminated encephalomyelitis. These findings demonstrate that lymphocytic cells reactive to myelin basic protein are present in the spinal fluid during active demyelinating disease; and that these cells may be more reactive than peripheral blood lymphocytes.

Acute Disease↗

Immunocytochemistry of cerebrospinal fluid.

In order to determine how best to study cells in cerebrospinal fluid (CSF) by immunocytochemical techniques, several crucial technical variables and five immunocytochemical methods were examined. Immunocytochemical studies could be performed on either cell suspensions or smears. The method using cell suspensions was more sensitive, producing less background staining, but requiring more cells than that using smears. Among the five methods examined, indirect immunoperoxidase (IP) and indirect immunoalkaline phosphatase (IAP) were comparable in sensitivity. The peroxidase-antiperoxidase (PAP), alkaline phosphatase-antialkaline phosphatase (APAAP) and avidin-biotin complex-immunoalkaline phosphatase (ABC-AP) methods were comparable in sensitivity and were more sensitive than either the IP or IAP technique. The peroxidase methods were plagued with problems related to endogenous enzyme activity and the ABC-AP method may exhibit undesirable background staining. Therefore, the IAP method should be used for cell suspensions and the APAAP for cells on smears. In CSF specimens with a small number of cells, immunocytochemical studies should be done on smears by the APAAP method. These conclusions are supported by our experience with CSF specimens from patients with reactive and neoplastic lymphocytoses.

Adult↗

Clinical utility of broth cultures of cerebrospinal fluid from patients at risk for shunt infections.

For patients with cerebrospinal fluid (CSF) shunts, culture of the CSF remains the most valuable tool in the evaluation of suspected shunt infections. To detect anaerobic Propionibacterium sp., a well-described cause of these infections, many clinical microbiology laboratories routinely employ a broth medium as an adjunct to solid media. The use of broth, however, creates a diagnostic dilemma since many contaminants also are isolated from broth cultures. Therefore, we retrospectively reviewed the records of 59 patients with CSF shunts in whom an organism was isolated from only broth cultures to assess their utility for the diagnosis of shunt infection. We found that no single clinical or laboratory parameter, including fever, leukocytosis, pleocytosis, or CSF protein and glucose, could reliably predict or exclude a shunt infection. Isolation of coagulase-negative staphylococci only in broth, in the absence of growth on solid media in concurrent or immediately preceding cultures, virtually always represented contamination. The isolation of Propionibacterium sp. from broth only usually represented contamination; however, infection could not be excluded without a repeated CSF culture, even in the absence of pleocytosis. We recommend that specific comments be appended to laboratory reports for isolates from CSF in broth only as an aid to the physician in interpreting the clinical importance of such isolates.

Bacteriological Techniques↗

Somatostatin cerebrospinal fluid levels in dementia.

Somatostatin levels were measured in cerebrospinal fluid of patients with Alzheimer's disease, multi-infarct dementia and normal pressure hydrocephalus and compared with levels from a normal control group. All pathological groups showed a statistically significant decrease of somatostatin with respect to the control group, but no significant differences were found amongst them. A negative correlation was found between the Mini Mental State Test and the somatostatin levels in Alzheimer's disease patients but not in the other groups. Our results confirm that the lower levels of somatostatin in cerebrospinal fluid are not specific to Alzheimer's disease and indicate that the decrease found in all the groups is probably the result of neuronal destruction or damage in the diseases examined.

Adult↗

Semiautomatic analysis of phase contrast magnetic resonance imaging of cerebrospinal fluid flow through the aqueduct of Sylvius.

OBJECTIVE: Quantification of the cerebrospinal fluid (CSF) flow through the aqueduct of Sylvius by means of magnetic resonance imaging (MRI) is subject to interobserver variability due to the region of interest (ROI) selection. Our objective is to develop a semiautomatic measurement method to achieve reproducible quantitative analysis of CSF flow rate and stroke volume. MATERIAL AND METHODS: MR examinations were performed using a 1.5 T scanner with a phase contrast sequence (velocity encoding [V(enc)] of 20 cm/s, FOV = 160, 3 mm slice thickness, image matrix size = 256x256, TR = 53 ms, TE = 11 ms, NSA = 2, flip angle = 15 degrees and 23 frames per cardiac cycle with peripheral retrospective pulse gating). Our method was developed using MATLAB R7. Errors introduced by background offset and possible aliased pixels were automatically detected and corrected if necessary in order to calculate the flow parameters that characterize CSF dynamics. The semiautomatic seed method reproducibility was evaluated and compared with the radius method by two observers analysing 21 healthy subjects. RESULTS: The measurements using the semiautomatic seed method reduced the interobservers variability (intra-class correlation [ICC] = 1.0 for stroke volume and for volumetric flow rate) versus the radius method (ICC = 0.46 for stroke volume and 0.65 for flow rate). Normal stroke volume (39.19 +/- 20.13 microl/cycle), flow rate (3.81 +/- 2.81 ml/min), maximal mean systolic velocity (5.27 +/- 1.3 cm/s) and maximal mean diastolic velocity (4.20 +/- 1.4 cm/s) were calculated with the half moon and aliasing corrected seed method. CONCLUSIONS: Semiautomatic measurements (seed method with half moon background and aliasing correction) allow a generalization of the calculus of flow parameters with great consistency and independency of the operator.

Algorithms↗

Cerebrospinal fluid constituents collected at the atlanto-occipital site of xylazine hydrochloride sedated, healthy 8-week-old Holstein calves.

Cerebrospinal fluid (CSF) collected at the atlanto-occipital site and serum were obtained from 10 male, 8-week-old, Holstein calves after sedation with xylazine hydrochloride. Glucose, creatine kinase, alkaline phosphatase, urea nitrogen, creatinine, sodium, potassium, chloride, calcium, phosphorus, total protein, and albumin were determined in serum and CSF. Optical characteristics, specific gravity, total red blood cell and nucleated cell counts and differentials were also evaluated in the CSF. Additionally, CSF protein electrophoresis and immunoglobulin concentrations were determined. Then, albumin quotients (AQ) were derived. Erythrocytes were observed in 9 of 10 CSF samples. Total nucleated cell counts ranged from 0-10 cells x 10(6)/L with a mean of 3 cells x 10(6)/L. Differential nucleated cell count in the CSF consisted primarily of lymphocytes/small mononuclear cells (57%), fewer monocytes/ large mononuclear cells (38%), and scant neutrophils (4%) and eosinophils (0.05%). The concentration of sodium (134 to 139 mEq/L) was similar to that of serum, but the concentration of potassium (2.8 to 3 mEq/L) was lower than that of serum. Creatine kinase activity (0 to 4 U/L) of CSF was markedly lower than serum activity. The CSF glucose concentration was approximately 80% of the serum value. Cerebrospinal fluid total protein concentration determined by electrophoresis ranged from 110 to 330 mg/L with a mean of 159 mg/L. Cerebrospinal fluid albumin ranged from 48 to 209 mg/L with a mean of 86 mg/L. In all CSF samples, radial immunodiffusion of unaltered CSF and concentrated CSF (four-fold concentration) revealed quantities undetectable by the present techniques in which the lowest standard values for IgG1, IgG, and IgM determinations was 70 mg/L and IgG2 was 30 mg/L. The albumin quotient ranged from 0.15 to 0.65 with a mean of 0.25. Based on the results of this study, CSF may be collected at the atlanto-occipital site safely and efficiently in calves, and reported values for CSF from adult cattle may not be suitable for evaluation of CSF collected from immature cattle.

Alkaline Phosphatase↗

Immunochemical analysis of some proteins in cerebrospinal fluid and serum of patients with ischemic strokes.

Immunochemical studies of gamma gamma-neuron specific enolase (NSE), parvalbumin (PV), S-100 protein (S-100) and acidic fibrillary glial protein (GFAP) were studied in the cerebrospinal fluid and blood serum in 7 patients with ischemic cerebral stroke, aged 57 to 81 years. Cerebrospinal fluid and the first blood sample were taken on the first or second day of the disease. Further blood samples were taken once a week till the end of patients hospitalization, ending by patients discharge or death. Immunochemical identification of proteins under study were performed with Western-blotting technique. It was found that all proteins studied were present in both cerebrospinal fluid and blood serum on the first two days of the disease in small quantities. The blood content of both NSE and PV increased significantly during the first week of the disease. Both proteins disappeared from the blood serum between the second and fourth disease weeks. S-100 protein and GFAP contents in the blood reached significantly high level within the time interval between second and fifth disease weeks, and remained at a relatively high level till patients' death. In all cases computed tomography study and/or brain autopsy revealed extensive ischemic foci localized within areas supplied by the middle cerebral artery. No clear-cut correlation between extensiveness of the ischemic cerebral damage and the content of the proteins studied in both cerebrospinal fluid and blood serum was found. However, our data indicate that serial studies of the above proteins in patients with ischemic stroke may be useful in monitoring the progress of the disease, and occasionally in the prognosis at least in some cases.

Aged↗

[Lysozyme concentration in the cerebrospinal fluid of children with epilepsy and subacute sclerosing encephalitis].

Lysozyme concentration in the cerebrospinal fluid was determined in 21 children with epilepsy and 22 children with subacute sclerosing panencephalitis. In most children in both groups the concentration of the enzyme was in the range of control values, but in the remaining children it was above the upper limit of these values. No correlation was demonstrated between lysozyme concentration and total protein level in the cerebrospinal fluid. The causes of raised values of cerebrospinal concentration in these children are discussed.

Acute Disease↗

[Pilot study of the relationship of free amino acids in serum and in the cerebrospinal fluid of horses].

In a blind study serum and cerebrospinal fluid (CSF) of control horses and of horses in hepatic coma after chronic food intoxication with Senecio alpinus were collected simultaneously and the composition of free amino acids was determined. The hepatic encephalopathy index in serum (less than 1.65) and in CSF (less than 1.11) of liver patients was distinctly less than to the control values in serum (greater than 2.42) and in CSF (greater than 1.49). The serum concentrations of glutamic acid in hepatic coma were elevated five-fold in comparison to the controls. An indication of ammonia decontamination was that nearly ten-fold higher values of glutamine were found in the cerebrospinal fluids of patients than in serum. In comparison to controls the serum levels of glutamine in horses with hepatic encephalopathy were decreased by the factor 0.7.

Amino Acids↗

Differential neurotrophin levels in cerebrospinal fluid and their changes during development in newborn rat.

Cerebrospinal fluid concentration of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin-3 (NT-3) was measured in normal developing rat from birth to postnatal day (PND) 21 by enzyme-linked immunosorbent assay. NGF levels were significantly higher than those of BDNF and NT-3 from PND 1-21. NGF levels decreased from PND 1-3 to PND 9. At PND 15 and 17, NGF levels peaked a second time and rapidly decreased to PND 21. BDNF peaked at PND 13-15, while NT-3 levels peaked at PND 7-9. Each of the three neurotrophins has its own characteristic pattern in changes in cerebrospinal fluid levels.

Aging↗