Combined proteomic and metabolomic analyses of cerebrospinal fluid from mice with ischemic stroke reveals the effects of a Buyang Huanwu decoction in neurodegenerative disease.

Ischemic stroke is one of the most common causes of death worldwide and is a major cause of acquired disability in adults. However, there is still a need for an effective drug for its treatment. Buyang Huanwu decoction (BHD), a traditional Chinese medicine (TCM) prescription, has long been used clinically to aid neurological recovery after stroke. To establish potential clinical indicators of BHD efficacy in stroke treatment and prognosis, we conducted a combined proteomic and metabolomic analysis of cerebrospinal fluid (CSF) samples in a mouse stroke model. CSF samples were obtained from male mice with acute ischemic stroke induced by middle cerebral ischemic/reperfusion (CI/R) injury, some of which were then treated with BHD. Label-free quantitative proteomics was conducted using nano-LC-MS/MS on an LTQ Orbitrap mass and metabolomic analysis was performed using nanoprobe NMR and UHPLC-QTOF-MS. The results showed that several proteins and metabolites were present at significantly different concentrations in the CSF samples from mice with CI/R alone and those treated with BHD. These belonged to pathways related to energy demand, inflammatory signaling, cytoskeletal regulation, Wnt signaling, and neuroprotection against neurodegenerative diseases. In conclusion, our in silico data suggest that BHD treatment is not only protective but can also ameliorate defects in pathways affected by neurological disorders. These data shed light on the mechanism whereby BHD may be effective in the treatment and prevention of stroke-related neurodegenerative disease.

Cerebrospinal Fluid Metabolomics After Natural Product Treatment in an Experimental Model of Cerebral Ischemia.

Cerebrospinal fluid (CSF) is an important biofluid for diagnosis of and research on neurological diseases. However, in-depth metabolomic profiling of CSF remains an analytical challenge due to the small volume of samples, particularly in small animal models. In this work, we report the application of a high-performance chemical isotope labeling (CIL) liquid chromatography-mass spectrometry (LC-MS) workflow for CSF metabolomics in Gastrodia elata and Uncaria rhynchophylla water extract (GUW)-treated experimental cerebral ischemia model of rat. The GUW is a commonly used Traditional Chinese Medicine (TCM) for hypertension and brain disease. This study investigated the amine- and phenol-containing biomarkers in the CSF metabolome. After GUW treatment for 7 days, the neurological deficit score was significantly improved with infarct volume reduction, while the integrity of brain histological structure was preserved. Over 1957 metabolites were quantified in CSF by dansylation LC-MS. The analysis of this comprehensive list of metabolites suggests that metabolites associated with oxidative stress, inflammatory response, and excitotoxicity change during GUW-induced alleviation of ischemic injury. This work is significant in that (1) it shows CIL LC-MS can be used for in-depth profiling of the CSF metabolome in experimental ischemic stroke and (2) identifies several potential molecular targets (that might mediate the central nervous system) and associate with pharmacodynamic effects of some frequently used TCMs.

CSF orexin levels of Parkinson's disease, dementia with Lewy bodies, progressive supranuclear palsy and corticobasal degeneration.

Excessive daytime sleepiness has been widely accepted as a common problem not only in Parkinson's disease (PD) but also in other related disorders. Lowered excretion of orexin A (hypocretin 1) into the cerebrospinal fluid (CSF) is known to play a pathological role in narcolepsy and secondary hypersomnia due to hypothalamic dysfunction. Although the levels of CSF orexin in PD have been previously examined, the results have been controversial, and no systematic investigation of CSF orexin excretion has been conducted on PD related disorders. In this study, orexin was measured in CSF collected by lumbar puncture in 62 patients with PD, 13 patients with dementia with Lewy bodies (DLB), 16 patients with progressive supranuclear palsy (PSP), and 7 patients with corticobasal degeneration (CBD). Levels of CSF orexin (mean+/-SD pg/ml) were 302+/-38 in PD, 297+/-48 in DLB, 258+/-37 in PSP, 246+/-90 in CBD. The occurrence of low orexin levels (

[A 26-year-old woman of interval form of acute carbon monoxide intoxication with cerebrospinal fluid abnormalities].

A 26-year-old woman was admitted to our hospital for the treatment of hyperbaric oxygen therapy to acute carbon monoxide intoxication. The consciousness disturbance improved and she was discharged after 23 times of the hyperbaric oxygen therapy. However, she was readmitted because of dementia and urinary incontinence after 22 days. Diffusion-weighted images showed bright high signal intensities in the periventicular white matter and corpus callosum. The condition was considered to be an interval form of carbon monoxide intoxication. She was treated by 38 times of the hyperbaric oxygen therapy with cytochrome C and fully recovered. MRI images and cerebrospinal fluid abnormality (high protein content and IgG index) became normalized somewhat later than the improvement of the symptoms. By an investigation utilizing diffusion-weighted images, we thought that not only the demyelination which mentioned formerly, but the vasogenic edema was involving in the mechanism of these high signal intensities in the periventicular white matter of the interval form. And in the range which we searched, this is the first report which mentioned the abnormal findings of cerebrospinal fluid in an interval form of carbon monoxide intoxication. So we believe this case is very important for telling us suspected the mechanism and some indications about the treatment of an interval form.

Characteristic transfer of colostrum-derived biologically active substances into cerebrospinal fluid via blood in natural suckling neonatal pigs.

The characteristic transfer of colostral components into cerebrospinal fluid (CSF) via serum after natural suckling has been studied by sodium dodecyl sulphate (SDS) electrophoresis, two-dimensional electrophoresis, immunoblot and enzyme linked immunosorbent assay methods in non-suckling pigs. Total protein concentrations in the serum increased immediately after first suckling, reached a peak value at 12 h, corresponding to a 2.3-fold compared with pre-suckling level. The protein concentration in CSF also increased and reached peak value at 6 h corresponding to 1.6-fold compared with presuckling level. IgG in serum not detected before suckling, increased steeply after suckling, IgG, IgM and IgA transported into the serum were observed in completely intact form by immunoblot method. The IgG transported into serum was quickly transferred into CSF after natural suckling in contrast to the case of bovine IgG. Serum concentration of transferrin was maintained at high level before suckling and was not changed by suckling. Transferrin also detected in CSF was not changed by suckling. Bovine lactoferrin (Lf) administered into the intestinal lumen was transported into serum (0.01%) and also detected in CSF after 6 h as undegraded form (3.1%). Thus, homologous IgG and bovine Lf are transported into CSF, suggesting that the transport of macromolecules into CSF is selective in neonatal pigs.

Transport of colostral macromolecules into the cerebrospinal fluid via plasma in newborn calves.

The objective of this study was to investigate the transfer of bovine colostral macromolecules especially the lactoferrin (Lf), transferrin (Tf), immunoglobulin G (IgG), and epidermal growth factor (EGF) from the gastrointestinal tract to the cerebrospinal fluid (CSF) via systemic circulation in newborn calves. Cannulae were placed into the jugular vein and cisterna magna to collect blood and CSF, respectively at various time points. The colostrum, plasma, and CSF were analyzed by ELISA, SDS-PAGE, two-dimensional PAGE, and Western blotting. The concentration of total protein, Lf, Tf, and IgG in plasma averaged 47 mg, 204 ng, 101 microg and 15 microg/ml before colostrum feeding and increased to the peak values of 64 mg, 2413 ng, 820 microg, and 4608 microg/ml 8 h after feeding, respectively. Before colostral feeding CSF, total protein, Lf, Tf, and IgG averaged 0.44 mg, 10.3 ng, 0.31 microg, and 0.11 microg/ml, but peak values after feeding averaged 2.0 mg, 173 ng, 71 microg and 72 microg/ml after 10 h, respectively. Immunologically, six EGF-positive protein bands were detected in colostrum as well as in three bands higher density in plasma and CSF after colostral feeding. This study revealed that the colostral macromolecules were not only absorbed into the systemic circulation, but also some of them including Lf, Tf, IgG, and EGF-like proteins were transported into the CSF in a time-dependent manner through blood-CSF or blood-brain barrier of the newborn calves.

Characteristic transfer of colostral components into cerebrospinal fluid via serum in neonatal pigs.

In order to evaluate the possibility of modification of brain function by colostral suckling, the characteristic transfer of colostral components into serum and cerebrospinal fluid (CSF) has been studied by SDS electrophoresis, immunoblot and ELISA methods in nonsuckling pigs. Total protein concentrations in the serum increased immediately after oral administration of bovine colostrum, reaching a peak value (7.0 +/- 0.7 g/dl) at 24 h after administration, corresponding to a 3-fold increase compared to preinfusion levels. IgG and other macromolecular components (MW 19, 000-58,000) were recognized in serum by electrophoretic and ELISA analysis. Total protein concentrations in the CSF collected from the cisterna magna also increased steeply after colostral administration, reaching a maximal value (54.1 +/- 5.0 mg/dl) at 4 h, corresponding to a 4-fold increase compared to preinfusion levels. Two colostral components (MW 19,000 and 31,000) in serum were confirmed to be present in the CSF by electrophoresis. The component of MW 19,000 was identified by immunoblot as beta-lactoglobulin. IgG in serum transferred from colostrum could not be detected in the CSF by ELISA. Lactoferrin administered into the intestine was also detected in the CSF via serum. These results indicate that some components of colostrum can be transported into the CSF via the serum, suggesting the possibility of modification of immature brain functions by colostral suckling in neonatal pigs.

Treatment of bacterial meningitis with ceftizoxime.

Ceftizoxime was evaluated in the treatment of 18 patients (6 adults and 12 children) with bacterial meningitis. In seven patients Haemophilus influenzae was the causative agent, in three Neisseria meningitidis, in five Streptococcus pneumoniae, and in one each alpha-streptococcus and Escherichia coli; one case was culture negative. Ceftizoxime was administered intravenously in doses of 200 mg/kg per day. Clinical response was appropriate in all patients with a mean time of defervescence of 3.7 days, and sterile cerebrospinal fluid was obtained from all patients at 24 to 36 h after initiation of therapy. The mean concentration of ceftizoxime in 46 cerebrospinal fluid samples obtained during therapy was 8.53 micrograms/ml (range, less than 0.5 to 29.0 micrograms/ml). Ceftizoxime concentrations in cerebrospinal fluid samples were ten- to several hundredfold the bactericidal concentrations of the pathogens isolated from the cerebrospinal fluid. Ceftizoxime penetrates the meninges well during acute infection and appears to be an excellent candidate antibiotic in the treatment of bacterial meningitis.

Pharmacology and cerebrospinal fluid penetration of moxalactam in children and dosage recommendations.

The pharmacokinetics and penetration into cerebrospinal fluid (CSF) of moxalactam were evaluated in 20 children with bacterial meningitis. After moxalactam was given iv as a loading dose of 100 mg/kg followed by two doses of 50 mg/kg every 6 hr, the mean serum concentrations were 205 micrograms/ml at the end of the last infusion and 11 micrograms/ml at 6 hr. The beta elimination half-life was 3.6 hr for children younger than nine months of age and 1.7 hr for children older than nine months of age. The mean CSF concentration of moxalactam was 3.8 micrograms/ml (range, 0.9-22.6 micrograms/ml). The mean concentration of moxalactam in CSF was 19.1% (range, 3.1%-97%) that in serum. Bactericidal titers were at least 1:32 in 12 CSF specimens. Concentrations of moxalactam in CSF correlated significantly (P less than 0.001) with concentrations of protein in CSF. Concentrations of moxalactam in CSF were greatly in excess of minimal inhibitory/bactericidal concentrations for Haemophilus influenzae type b. Dosages of moxalactam for the treatment of systemic infections due to H. influenzae type b in children are proposed.