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Comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC×GC-TOFMS) was applied in the quantification and identification of organic compounds in patient-matched human cerebrospinal fluid (CSF) and serum samples. Concentrations of 21 amino and hydroxyl acids varied from 0.04 to 77ng/μl in CSF and from 0.1 to 84ng/μl in serum. In total, 91 metabolites out of over 1200 detected were identified based on mass spectra and retention indices. The other metabolites were identified at the functional group level. The main metabolites detected in CSF were sugar and amino acid derivatives. The CSF and serum had clearly distinct metabolic profiles, with larger biological variation in the serum than in CSF. The GC×GC-TOFMS allowed detection and identification of several metabolites that have not been previously detected in CSF., (Copyright © 2013 Elsevier B.V. All rights reserved.)

A 29-year-old man diagnosed as having pulmonary sarcoidosis in 2008, and hypothyroidism secondary to thyroidectomy for Basedow's disease was admitted to our hospital with pustular psoriasis in November 2010. He experienced high fever (38°C) and headache in late October 2010. Gadolinium-enhanced T1-weighted image showed multiple micronodular lesions with leptomeningeal enhancement, mainly in the brainstem. Cerebrospinal fluid (CSF) analysis revealed pleocytosis, raised protein level and hypoglycorrhachia (7 mg/dl). The patient was also found to have osteonecrosis of the left femoral head. Antituberculous treatment and steroid pulse therapy were started, but produced no improvement of either the symptoms or the laboratory data. Finally, the patient was diagnosed as having meningeal disseminated sarcoidosis by meningeal biopsy in late March 2011. He was started on treatment with 60 mg prednisolone per day, which resulted in marked clinical improvement. It should be borne in mind that marked hypoglycorrhachia in the CSF can also be seen in meningeal disseminated sarcoidosis.

We investigated whether blood N-glycan changes can be used as a diagnostic biomarker for Alzheimer disease (AD). We used DNA sequencer-assisted, fluorophore-assisted carbohydrate electrophoresis (DSA-FACE) technology to assay N-glycans in sera from 79 autopsy-confirmed dementia patients and 149 healthy controls. One N-glycan (NA2F) was substantially decreased in AD patients but not in controls. Use of NA2F for discriminating AD between dementia patients and healthy controls showed a diagnostic accuracy of 85.7% +/- 2.8% with 92% specificity and 70% sensitivity. The decrease in the level of NA2F in AD patients compared to non-AD patients was more pronounced in females (p < 0.0001) than in males (p < 0.014). Use of NA2F to differentiate female AD from female non-AD patients reached a diagnostic accuracy of 90.7% +/- 4.8 %. Pearson correlation analysis showed that in female dementia patients, serum NA2F levels were significantly correlated with the cerebrospinal fluid (CSF) beta-amyloid peptide of 42 amino acids (Abeta(1-42)) and tau phosphorylated at threonine 181 (P-tau(181P)) levels, whereas in male dementia patients serum NA2F levels were significantly correlated only with CSF total tau protein (T-tau) level. Thus, we suggest that the serum N-glycan marker might be suitable for longitudinal and follow-up studies.

Between 1991-2006, nine patients below age 18 years, with a microbiologic documentation of Cryptococcus neoformans infection and no evidence of human immunodeficiency virus infection, were identified and treated at Chang Gung Children's Hospital. All exhibited central nervous system involvement. Seven patients were female (age range, 9-16 years; mean age, 13.7 years). Five patients (56%) manifested underlying diseases and were receiving either steroid or immunosuppressant treatment at time of disease onset. Eight patients presented with meningitis. Headache, vomiting, and focal neurologic signs were the most common presentations. Protein and sugar levels in cerebrospinal fluid were within normal range in seven cases, whereas India ink smear and cryptococcal antigen testing were positive in 87% (7/8) and 78% (7/9) of patients, respectively. With prompt antifungal therapy, all survived, but one presented the sequel of blindness. Cryptococcosis is uncommon in the nonhuman immunodeficiency virus-infected pediatric population. Clinicians should take into account a diagnosis of central nervous system cryptococcosis when children present with prolonged headache, vomiting, and focal neurologic signs. Indian ink stain and cryptococcal antigen testing of cerebrospinal fluid should be performed., (Copyright 2010 Elsevier Inc. All rights reserved.)

Background: There is no information regarding the possible role of cerebral substrates in the pathogenesis of neuronal injury in intracerebral haemorrhages (ICHs). Purposes of this prospective study were to clarify whether changes in substrates are the consequence of the initial brain damage in ICH and to elucidate the relationship among the biochemical mechanisms and clinical course of patients with ICH., Method: During a period of two years, patients (GCS < or =8) who had ICH secondary to an aneurysm (SAH), stroke (sICH), or trauma (tICH) and underwent ventriculostomy with ICP monitoring and/or underwent cranial surgery were randomly enrolled in this study. Extracellular concentrations of glutamate, aspartate, glycine, GABA, lactate, lactate/pyruvate ratio, and glucose in the CSF were measured by use of high-performance liquid chromatography (HPLC). The nitric oxide (NO) concentration in the CSF was analyzed by chemiluminescence., Findings: There were 75 patients (38 women and 37 men) with ICH included in this study. Twenty-one patients had SAH, 28 sICH, and 26 tICH. In tICH patients, there was a 30-fold increase in glutamate and a 10-fold in aspartate over reference values. The levels of glutamate, aspirate, GABA, lactate, glucose, and NO differed significantly among the three groups (p<0.001). There were no significant differences in glycine and L/P ratio among the groups. The initial GCS, the mean CPP and outcome six months after the insult were all significantly correlated with the concentration of substrates (p<0.01), both within groups and among the total sample. The CSF levels of glutamate lactate, NO and glucose correlated significantly with outcome (p<0.005)., Conclusions: This study confirms the correlation between the level of EAAs and the outcome of ICHs, suggesting that neurochemical monitoring of these substances may have a role in caring for patients.

Clusterin is suggested to be involved in the pathogenesis of Alzheimer's disease. Clusterin expression is increased in brain tissue in affected regions of Alzheimer patients, and intense clusterin staining is found in both senile plaques and in neuronal and glia cells. In contrast, the cerebrospinal fluid level of clusterin in Alzheimer patients has, thus far, been found unchanged. Clusterin is a glycosylated protein, and an alteration of its glycosylation in Alzheimer's disease might influence accurate quantification in cerebrospinal fluid through interference of antibody binding to the protein. Using enzymatic deglycosylation of clusterin isolated from cerebrospinal fluid, we found that the carbohydrates attached to clusterin were of the N-linked type and sialic acids. Based on this finding, cerebrospinal fluid samples from Alzheimer patients (n=99) and controls (n=39) were analysed. The samples were treated with peptide: N-glycanase F, cleaving off N-linked carbohydrates, and clusterin was quantified before and after deglycosylation using a new sandwich enzyme-linked immunosorbent assay. Clusterin was significantly increased in Alzheimer patients, in both native (7.17+/-2.43 AU versus 5.73+/-2.09 AU; p=0.002), and deglycosylated samples (12.19+/-5.00 AU versus 9.68+/-4.38 AU; p=0.004). Deglycosylation led to increased measured levels of clusterin by 70% (p<0.001) in Alzheimer patients and 67% (p<0.001) in controls. These findings indicate that glycosylation of proteins may interfere with their quantification. The results show that clusterin is significantly increased in cerebrospinal fluid from Alzheimer patients as a group, supporting that clusterin might be involved in the pathogenesis of Alzheimer's disease. However, the individual clusterin levels overlap between the two groups, and thus cerebrospinal fluid clusterin measurement is not suitable as a biochemical marker in the diagnosis of Alzheimer's disease.

The present article describes the first patient with a deficiency of ribose-5-phosphate isomerase (RPI) (Enzyme Commission number 5.3.1.6) who presented with leukoencephalopathy and peripheral neuropathy. Proton magnetic resonance spectroscopy of the brain revealed highly elevated levels of the polyols ribitol and D-arabitol, which were subsequently also found in high concentrations in body fluids. Deficient activity of RPI, one of the pentose-phosphate-pathway (PPP) enzymes, was demonstrated in fibroblasts. RPI gene-sequence analysis revealed a frameshift and a missense mutation. Recently, we described a patient with liver cirrhosis and abnormal polyol levels in body fluids, related to a deficiency of transaldolase, another enzyme in the PPP. RPI is the second known inborn error in the reversible phase of the PPP, confirming that defects in pentose and polyol metabolism constitute a new area of inborn metabolic disorders.

Extensive assignments of resonances in the 600 MHz 1H-NMR spectra of cerebrospinal fluid are reported. These have been achieved by the measurement of a combination of two-dimensional experiments comprising homonuclear J-resolved, COSY45, and double-quantum filtered COSY (DQCOSY) spectra. By these means the previous total of 18 endogenous metabolites, of which in general only selected resonances have been assigned, has been augmented to 46 molecules including all of the resonances of both alpha- and beta-anomers of glucose. With only a few exceptions all resonances have been assigned for all of the metabolites. In addition, the effect of freeze-drying on the 600 MHz 1H-NMR spectrum of human cerebrospinal fluid (CSF) is presented using both lyophilization with reconstitution into either H2O or D2O. Freeze-drying and reconstitution into H2O causes a significant sharpening of many small molecule resonances, including notably those of glutamate and glutamine as well as other amino acids and in addition causes the loss of volatile components, principally acetone. Further exchange of the H2O solvent by D2O causes no additional changes in the spectra.

Six non-glucose polyols--mannose, fructose, 1-deoxyglucose, mannitol, glucitol, and inositol--were identified and evaluated in human serum and cerebrospinal fluid by gas-liquid chromatography and by gas-liquid chromatography/mass spectrometry. Concentrations of fructose, mannose, and inositol in the serum of healthy persons or children without metabolic diseases varied with age, as already reported for 1-deoxyglucose. Fructose, inositol, and glucitol concentrations in cerebrospinal fluid significantly exceeded those in serum. The method described here for determining polyols and for evaluating polyol patterns in serum, as well as the resulting data on children and healthy subjects, should be useful in investigations of the clinical and physiological significance of polyols.

We used gas chromatography in conjunction with flame ionization detection to quantitate nine polyols and aldo and keto sugars (as silyl derivatives) in human plasma and cerebrospinal fluid (CSF). Rhamnose, not found in CSF or plasma, was used as an internal standard with a lower limit of quantitation of 0.4 mg/l. CSF polyol and sugar concentrations (mean +/- S.D.) in fourteen healthy subjects (age range 27.1-85.9 years) were: anhydroglucitol, 19.9 +/- 5.3 mg/l; arabitol, 4.8 +/- 0.9 mg/l; erythritol, 2.4 +/- 0.5 mg/l; myoinositol, 28.6 +/- 8.3 mg/l; ribitol, 1.6 +/- 0.1 mg/l; fructose, 25.5 +/- 11.1 mg/l; glucose, 587 +/- 70 mg/l; glucitol, 7.7 +/- 1.5 mg/l; and mannose, 10.6 +/- 2.4 mg/l. The respective plasma concentrations were 30.6 +/- 11.5, less than 0.4, 0.4 +/- 0.2, 6.3 +/- 2.6, less than 0.4, 23.4 +/- 21.4, 897 +/- 214, less than 0.4 and 13.7 +/- 6.3 mg/l. Polyol CSF-to-plasma concentration ratios greater than 2 were observed for myoinositol, erythritol, arabitol, glucitol and ribitol, indicative of active accumulation or synthesis of these polyols within the central nervous system.

A case of measles meningoencephalitis is described. Measles virus was isolated from the cerebrospinal fluid cells, in which virus antigen was also detected by indirect immunofluorescence.

A study of measles was undertaken from the case histories of 3 093 children under 15 years of age, treated in the children's clinics of Hamburg between 1960 to 1973. 562 children were suffering from pneumonia (18,2%), 230 had otitis (7,4%) and 229 (7,4%) showed neural affections (143 cases with fits and 86 patients with encephalopathy or encephalitis), Various parameters were analysed: age, sex, seasonal distribution, fatality, age disposition to convulsions and measles encephalitis, "incubation period" of neural affections, sequelae. Permanent sequelae due to measles encephalitis were seen in 14% of the cases. A plea is made for an universal immunisation against measles.

Fluid from the notochordal canal of the coelacanth, Latimeria chalumnae, was analyzed for major inorganic and organic constituents and compared with blood serum from the same fish. Significantly or suggestively lower levels of sodium, magnesium, calcium, bicarbonate, sulfate, total carbohydrates, glucose, lactate, cholesterol, bound phosphate and total proteins were found in notochordal fluid than in serum, whereas potassium, chloride, urea, trimethylamine oxide, and total free amino acids were higher and inorganic phosphorus essentially identical. Osmolarity of notochordal fluid (1058 mOsm) exceeds that of serum (942 mOsm). A whitish precipitate in the fluid consisted of a matrix of fibers 100 A in diameter and of indefinite length. It resembled a sialoglycoprotein in composition and was stabilized by disulfide bonds. The fluid contained cellular debris.

The sugar content in the liquor of rabbits exposed to accelerations ranging from several minutes to several hours was studied. The animals were accelerated at 3 g in a centrifuge with a radius of 55 cm and a velocity of 70 rpm, in the Khilov's swing and in a rotating device simulating angular accelerations. The liquor was sampled by the suboccipital puncture and the sugar content was measured according to Hagedorn-Jensen. A statistically significant increase in the sugar content was noted after every exposure to accelerations of alternating signs than with accelerations of a constant value. The experiments on labyrinth-ectomized animals showed a certain effect of the vestibular analyzer on the carbohydrate metabolism. However, when the equilibrium function reached compensation, the sugar content in the liquor returned to the pretest level. Therefore, this reaction involves not only the vestibular system but also the optic, proprio- and interoceptive afferent systems. A comparison of the intracysternal and intramuscular injections of epinephrine points to the central (in contrast to the hematogenic) nature of an increase in the sugar content during the first minutes of acceleration. The electric stimulation of the anterior and posterior hypothalamus by means of implanted electrodes produced the same elevation of the liquor sugar as accelerations. It can be postulated that afferent impulses from the analyzer systems reach the hypothalamic centers, thus exciting the autonomic system and, first of all, the sympathic system.

The feasibility of gas-liquid chromatographic analysis of spinal fluid from dogs with experimental meningitis and from patients with meningitis was evaluated. Normal canine and human spinal fluid contained consistent levels of palmitic and stearic acids, glucose, and trace amounts of glycerol. Pneumococcal meningitis in dogs and humans was characterized by a complex fatty-acid pattern in spinal fluid similar to that seen in pneumococcal cells. Spinal fluid from dogs with Haemophilus influenzae meningitis yielded several saturated fatty acids as well as rhamnose, xylose, and glucosamine; four of five spinal fluid specimens from patients with H. influenzae meningitis gave similar results. Fatty-acid profiles of spinal fluid from dogs with staphylococcal meningitis were complex, with branched-chain and straight-chain fatty acids. Spinal fluid specimens from Neisseria meningitidis-infected dogs yielded 16- and 18-carbon fatty acids and sialic acid. Gas-liquid chromatographic analysis of spinal fluid proved valuable as a tool for identifying bacterial infections and differentiating between common spinal fluid pathogens.

A toxic syndrome characterized by fever, headache, and vomiting, lasting 2-5 days, occurred in 61% of 39 children with acute leukemia in complete remission, receiving central nervous system prophylaxis with intrathecal methotrexate, and in 14% of 34 children receiving the same plus cranial radiation. The syndrome was accompanied by pleocytosis with lymphocytes, monocytoid cells, and neutrophils. There was evidence of cumulative Mtx toxicity, since the toxic syndrome occurred mostly after the third and fourth dose and did not recur with longer intervals between doses. The incidence of the syndrome was significantly reduced by the use of Elliott's B solution as Mtx diluent, rather than water or normal saline. The occurrence of pleocytosis and toxic clinical syndrome was also significantly reduced in patients receiving concomitant cranial radiation, probably due to the lympholytic action of radiotherapy and the depressed cellular response of irradiated tissues. The use of Elliott's B solution as diluent for IT Mtx and an appropriate interval between Mtx doses are suggested for prevention of this toxic syndrome.

A highly reproducible procedure was developed for gas-liquid chromatographic analysis of trimethylsilyl derivatives of normal human cerebrospinal fluid. Fourteen normal human cerebrospinal fluid samples tested with this procedure contained alpha- and beta-glucose as well as isomers of two unidentified sugars. Chromatographic changes in three cases of meningeal inflammation (two cryptococcosis and one thalamic astrocytoma) were limited to decreased concentrations of all sugars. In one case of early meningitis, the concentrations of the unknown sugars decreased before glucose. Now that a reproducible chromatogram of the trimethylsilyl derivatives of normal human cerebrospinal fluid has been established, more samples of abnormal cerebrospinal fluid should be prepared by these methods and examined by gas-liquid chromatography. It may be possible to identify unique products of infectious agents which will permit rapid diagnosis of central nervous system infection.

Gas-liquid chromatographic procedures for measuring sugar and polyol concentrations in biological fluids are reviewed. Such methods require the preparations of derivatives such as methyl ethers, trimethylsilyl ethers or acetyl esters. Prior to derivatisation samples must be deproteinised and dried. Complex mixtures of sugars and sugar alcohols may be resolved. Quantitative analyses are precise, sensitive and linear. If internal standardisation is used recoveries approaching 100% are obtained.

A development of a technique for estimating sugar in blood, cerebrospinal fluid, etc., is described, using 3:5-dinitrosalicylic acid (D.N.S.A.) originally introduced by Sumner (1921). Results can be obtained in less than 10 minutes if required. The method is well suited to the estimation of random blood sugars and the handling of diabetic clinic requirements in hospital laboratories. The reagents are cheap, stable, and easily prepared. The results are very close to true glucose values in blood and cerebrospinal fluid. The technique has justified its existence in a busy clinical laboratory on the grounds of simplicity and rapidity, and is sufficiently precise for all ordinary work.