Content In Cerebrospinal Fluid Research Articles

Effect of Yishen Qiqiao Fang on dopamine content in serum and cerebrospinal fluid of patients in persistent vegetative statec*

Background It has been demonstrated that the attack of persistent vegetative state is associated with the damaged neuron, which produces dopamine, and nervous pathway, the reduce of dopamine or malfunction of dopamine. Objective To observe the effect of self-made traditional Chinese medicine (TCM) Yishen Qiqiao Fang, which has the functions of supplementing qi and nourishing blood, resolving phlegm by promoting blood circulation, restoring consciousness and inducing resuscitation, on the contents of dopamine in serum and cerebrospinal fluid of patients in persistent vegetative state. Design An open randomized controlled clinical trial. Settings Nanjing University of Traditional Chinese Medicine, Nanjing Zijin Hospital. Participants Thirty-eight inpatients of persistent vegetative state were selected from the Department of Neurology, Nanjing Zijin Hospital from August 2005 to November 2006. The patients were diagnosed according to the diagnostic standards set by the summary of a meeting for specialists in Nanjing. Informed contents were obtained from their relatives. According to the order of admission, the enrolled patients were divided into control group ( n =20) and TCM treated group ( n =20). Methods In the control group, the patients were treated with routine treatments for the symptoms. In the TCM treated group, the patients were treated with Yishen Qiqiao Fang besides the same treatments in the control group. TCM dispensing granules: each bag of mongolian milkvetch root, Chinese angelica and peach seed equaled to 10 g crude drug respectively; each bag of grassleaf sweetflag rhizome and dahurian angelica root equaled to 6 g crude drug respectively; Musk 0.05 g. The daily dosage for adults: 4 bags of mongolian milkvetch root, 2 bags of Chinese angelica, 0.05 g musk, 1 bag of peach seed, 2 bags of grassleaf sweetflag rhizome and 2 bags of dahurian angelica root, which should be given though nasal feeding or gastrostogavage before breakfast and supper every day, the course lasted for 2 months. The contents of dopamine in serum and cerebrospinal fluid were determined with enzyme immunoassay analyzer before and after treatment in both groups. Main outcome measures Contents of dopamine in serum and cerebrospinal fluid before and after treatment in both groups. Results Totally 38 patients of persistent vegetative state were enrolled, 33 of them were involved in the final analysis of results, and the other 5 missed. The contents of dopamine in serum and cerebrospinal fluid had no obvious differences between the two groups both before and after treatment ( P > 0.05). In the control group, the contents of dopamine in serum and cerebrospinal fluid had no obvious differences before and after treatment ( P > 0.05). In the TCM treated group, the contents of dopamine in serum and cerebrospinal fluid after treatment were obviously higher than those before treatment ( t =2.174, 2.485, P < 0.05). Conclusion Yishen Qiqiao Fang can significantly increase the contents of dopamine in serum and cerebrospinal fluid of patients in persistent vegetative state, which may be one of the mechanisms for ameliorating persistent vegetative state.

Blood-Brain Barrier Disruption Associated with Topiramate-Induced Angle-Closure Glaucoma of Acute Onset

Topiramate (Topamax(R)) is an anti-epileptic drug of the sulfamate group used secondarily for bipolar disease. One week after initiation of topiramate treatment for a bipolar disorder, a 57-year-old man presented with blurred vision. Clinical examination revealed a bilateral conjunctivitis, areflexic mydriasis, severe anterior chamber shallowing, with a myopic shift and vitritis. A spinal tap revealed an increased protein content of 1581 mg/L on cerebrospinal fluid (CSF) analysis, being compatible with a rupture of the blood-brain barrier (BBB). UBM exposed bilateral ciliochoroidal effusions with secondary angle-closure. Topiramate was promptly discontinued, whereas visual acuity, intraocular pressure (IOP), and anterior and posterior segments anatomy normalized within 1 week. One month later, bilateral iris atrophy was present. The presence of BBB disruption with increased protein content in CSF with simultaneous blood ocular barrier breakdown may suggest a common inflammatory mechanism.

Treatment of Neoplastic Meningitis With Intrathecal 9-Nitro-camptothecin

The topoisomerase I inhibitor, 9-nitro-camptothecin (9NC), is highly tumoricidal against glioblastoma multiforme (GBM) in vitro. However, systemic administration of 9NC has not shown the expected efficacy in clinical trials. This failure may be due to the rapid hydrolysis of 9NC in plasma from the active form to the inactive and myelosuppressive form in the presence of human albumin at physiologic pH. Concurrent treatment with anticonvulsants and dexamethasone, drugs indispensable for the supportive therapy of patients with GBM, has also been shown to decrease plasma concentrations of these drugs. Intrathecal drug delivery circumvents the blood-brain barrier and minimizes systemic toxicity. Intrathecal delivery of 9NC may also have the more specific advantage of significantly reducing the hydrolysis of 9NC that occurs after systemic delivery due to the more favorable pH and reduced albumin content in cerebrospinal fluid. The present study evaluated the toxicity and efficacy of intrathecal delivery of 9NC in an athymic rat model of neoplastic meningitis. Toxicity tests showed that 0.3 micromol (5000 microM), 0.03 micromol (500 microM), 0.003 micromol (50 microM), or 0.0003 micromol (5 microM) of 9NC administered intrathecally to the athymic rats caused no evidence of clinical or histological toxicity. Intrathecal administration of 0.3 micromol (5000 microM) of 9NC twice a week for three doses to athymic rats with neoplastic meningitis induced by the GBM cell line, U87MGDeltaEGFR, resulted in a 26% increase of median survival compared to the control group (p < 0.005). These results suggest that intrathecal treatment with 9NC may be useful for patients with GBM neoplastic meningitis.

Oxidative stress and metal content in blood and cerebrospinal fluid of amyotrophic lateral sclerosis patients with and without a Cu, Zn-superoxide dismutase mutation

Hydroxyl radical, ascorbate free radical, superoxide dismutase (SOD) activities, Cu,Zn-SOD protein, Mn-SOD protein, 8-hydroxy-2' -deoxyguanosine (8-OHdG) and metals were compared in red blood cells (RBC), plasma and/or cerebrospinal fluid (CSF) between patients with sporadic amyotrophic lateral sclerosis (SALS), familial ALS (FALS) showing the Leu126Ser mutation in the Cu, Zn-SOD gene and controls. In patients with FALS or SALS, concentrations of hydroxyl radical in blood and ascorbate free radical and 8-OHdG in CSF were higher than control group values, while SOD activities in RBC and CSF were lower. In contrast, Cu, Zn-SOD protein concentrations in RBC were low only in FALS patients. Concentrations of Cu in CSF of SALS patients were higher than in controls. Thus, the pathogenesis of increased oxidative stress differs between SALS patients and FALS patients with a mutant Leu126Ser SOD1 gene.

Methylmercury increases S100B content in rat cerebrospinal fluid

S100B, a calcium binding protein physiologically produced and released by astrocytes, has been used as a peripheral marker of brain damage. Here, we investigated the effects of subcutaneous injections of methylmercury chloride (MeHg–5 mg/kg), an environmental neurotoxicant, on S100B protein content in cerebrospinal fluid (CSF) of adult rats. In addition, the performance of animals in an open field (number of squares crossing and rearings) was also analyzed in order to obtain a possible link between alteration in S100B protein content in CSF and parameters related to neurological injury. MeHg treatment increased serum mercury and S100B protein levels in the CSF. A decrease in the numbers of crossings and rearings was observed in MeHg-treated animals when compared to control group, which suggests a possible neurological injury. The present data show, for the first time, increased S100B levels in CSF after exposure to a neurotoxic metal. Authors discuss the possibility of astrocytic involvement in MeHg-induced neurotoxicity.

Cerebrospinal Fluid S100B Increases Reversibly in Neonates of Methyl Mercury-Intoxicated Pregnant Rats

Methylmercury (MeHg), an organic methylated form of mercury, is one of the most hazardous environmental pollutants. MeHg is a potent neurotoxin, particularly during brain development. Neurotoxicity-induced by MeHg in prenatal age can cause mental disorders, cerebral palsy and seizures. We investigated cerebrospinal fluid (CSF) and brain tissue contents of S100B, a calcium binding protein produced and secreted by astrocytes, which has trophic and toxic activity on neurons depending on concentration. Pregnant rats were exposed to MeHg (5 mg/kg per day, on the 12th, 13th and 14th days of pregnancy). CSF and brain tissue (hippocampus, cerebral cortex and cerebellum) were obtained from neonate rats on 1, 15 and 30 days postnatal. MeHg accumulation was measured in brain tissue after birth and on the 30th postnatal day. An increase of CSF S100B was observed on the 15th, but not on the 30th postnatal day. Hippocampal tissue demonstrated increased S100B (and reduction in glial fibrillary acidic protein) immediately after birth, but not later. No changes in the S100B content were observed in cerebellum and cerebral cortex. No changes were observed in the spatial learning of these rats at adult age. These specific and reversible changes in the hippocampus could be related to the cognitive and epileptic disorders attributed to MeHg. Our results further indicate the glial involvement in the MeHg-induced neurotoxicity. The increment of CSF S100B in neonates exposed to MeHg reinforces the view that increased S100B is related to damage in the nervous system and that S100B could be a marker for MeHg-neurotoxicity. Although the cellular mechanism related to MeHg-induced increase in S100B content in CSF remains unknown, our results suggest the use of S100B as a peripheral marker of brain damage induced by MeHg.

Cystatin C as a cerebrospinal fluid biomarker for pain in humans

Through a process of subtraction cloning and differential hybridization, we previously identified several new genes whose expression was induced by peripheral inflammation. One of these coded for cystatin C, a secreted cysteine protease inhibitor in the cystatin superfamily. We hypothesized that, concurrent with increased expression in dorsal horn, increased secretion would elevate the cystatin C content in cerebrospinal fluid (CSF) during active pain states. Alterations were assessed by immunoassay and by surface enhanced laser desorption ionization (SELDI) mass spectrometry with either reverse phase or immobilized anti-cystatin C antibody surfaces using CSF from ten age-matched obstetrical patients at term. Five control subjects were scheduled for an elective caesarian section and were not in pain. Another five subjects were in labor for 8.9±1 h and were in severe pain as assessed with a visual analog scale and the McGill short form questionnaire. The level of cystatin C as measured by immunoassay in the non-pain patients was 2.77±0.75 μg/ml and in the pain patients 5.36±0.92 μg/ml ( P<0.02). The elevation occurred without significant change in total CSF protein or β-endorphin content. The cystatin C increase also was detectable by SELDI with either raw CSF or after antibody capture. These data are consistent with our previous animal study and the idea that persistent pain induces the synthesis and release of cystatin C in dorsal spinal cord, the surplus of which overflows into the CSF.

Developmental changes in S100B content in brain tissue, cerebrospinal fluid, and astrocyte cultures of rats.

1. We investigated the content of S100B protein by ELISA in three brain regions (hippocampus, cerebral cortex, and cerebellum) and in cerebrospinal fluid of rats during postnatal development as well as the content and secretion of S100B in pre- and postconfluent primary astrocyte cultures. 2. An accumulation of S100B occurred in all brain regions with similar ontogenetic pattern between second and fourth postnatal weeks. However, we observed a decrease in the cerebrospinal fluid S100B after the critical period for synaptogenesis in rodents. 3. A similar profile of cell accumulation and decrease in basal secretion was also observed during aging of astrocyte cultures. 4. These data contribute to the proposal that S100B is an important glial-derived protein during brain development and that changes in extracellular levels of S100B may be related to glial proliferation and synaptogenesis.

Endogenous Proteins Controlling Amyloid β-Peptide Polymerization: POSSIBLE IMPLICATIONS FOR β-AMYLOID FORMATION IN THE CENTRAL NERVOUS SYSTEM AND IN PERIPHERAL TISSUES

We report that certain plasma proteins, at physiological concentrations, are potent inhibitors of amyloid beta-peptide (Abeta) polymerization. These proteins are also present in cerebrospinal fluid, but at low concentrations having little or no effect on Abeta. Thirteen proteins representing more than 90% of the protein content in plasma and cerebrospinal fluid were studied. Quantitatively, albumin was the most important protein, representing 60% of the total amyloid inhibitory activity, followed by alpha1-antitrypsin and immunoglobulins A and G. Albumin suppressed amyloid formation by binding to the oligomeric or polymeric Abeta, blocking a further addition of peptide. This effect was also observed when the incorporation of labeled Abeta into genuine beta-amyloid in tissue section was studied. The Abeta and the anti-diabetic drug tolbutamide apparently bind to the same site on albumin. Tolbutamide displaces Abeta from albumin, increasing its free concentration and enhancing amyloid formation. The present results suggest that several endogenous proteins are negative regulators of amyloid formation. Plasma contains at least 300 times more amyloid inhibitory activity than cerebrospinal fluid. These findings may provide one explanation as to why beta-amyloid deposits are not found in peripheral tissues but are only found in the central nervous system. Moreover, the data suggest that some drugs that display an affinity for albumin may enhance beta-amyloid formation and promote the development of Alzheimer's disease.

Organophosphate-induced brain injuries: delayed apoptosis mediated by nitric oxide

The features of organophosphate-induced brain injuries were investigated. Rats were poisoned intraperitoneally with 9 mg/kg (1.8 LD 50) of diisopropylfluorophosphate. Pyridostigmine bromide (0.1 mg/kg) and atropine methylnitrate (20 mg/kg), which are centrally inactive, were pre-treated intramuscularly to reduce the mortality and eliminate peripheral signs. Diisopropylfluorophosphate induced severe limbic seizures, and early necrotic and delayed apoptotic brain injuries. The necrotic brain injury was observed to be maximal as early as 1 h after diisopropylfluorophosphate treatment predominently in hippocampus and piriform/entorhinal cortices, showing a spongiform change (malacia) of neuropils in severe cases. In contrast, typical apoptotic (TUNEL-positive) cells started to appear at 12 h in thalamus, and a mixed type in amygdala. Separately, nitrite/nitrate content in cerebrospinal fluid was found to significantly increase after 2 h, reaching a maximal level at 6 h. Pre-treatment with l- N G-nitroarginine, an inhibitor of nitric oxide synthase, reduced nitrite/nitrate content and, noteworthy, attenuated only apoptotic brain injury in all four brain regions without affecting seizure intensity and necrotic injury. Taken together, the delayed apoptotic injury of brain induced by diisopropylfluorophosphate poisoning in rats might be mediated in part through nitric oxide production.

Effect of ligustrazine on nitric oxide contents in cerebrospinal fluid and plasma of patients with cerebral infarction

To probe into the significance of Nitric Oxide (NO) in ischemic cerebral damage and effect of ligustrazine on it. The NO contents in cerebrospinal fluid (CSF) and plasma of 20 controls and 62 patients with arteriosclerotic thrombotic cerebral infarction (ligustrazine group and common treatment group) were determined with Griess method before and after treatment. The NO content in CSF before treatment was higher in severe type, it was higher than that in moderate type, and than that in mild type, again higher than that in control group (all P < 0.05) and was positively correlated with the size of infarction (P < 0.01). There was no significant difference between the plasma NO content of patients and that of control group (P > 0.05), but there was a trend that plasma NO content decreased as the patient's condition worsened. After treatment, the curative effect and plasma NO content of ligustrazine group were both markedly higher than those of common treatment group (P < 0.05). There were no significant differences between CSF NO contents of the two groups, respectively, and that of control group (P > 0.05). Excessive NO produced in brain tissue and insufficient plasma NO participate the course of ischemic brain damage. Ligustrazine could increase the contents of plasma NO selectively.

Nitro-L-arginine augments the endothelin-1 content of cerebrospinal fluid induced by cerebral ischemia

The effect of nitro-L-arginine (NLA), inhibitor of NO synthase, on ET-1 content in cerebrospinal fluid (CSF) and on the vascular system was investigated in global ischemia/reperfusion of Mongolian gerbils. The results indicate that NLA induced a prolonged (2–3-fold) increase of ET-1 concentration above that seen in the CSF of untreated animals during ischemia/reperfusion. Both the transient and prolonged rise of ET-1 content observed in the CSF coincided with the reduction in the cerebral blood flow seen in untreated and NLA-treated gerbils, respectively, at the time of reperfusion.

Macrophage inflammatory protein-1α in the cerebrospinal fluid of patients with multiple sclerosis and other inflammatory neurological diseases

The level of macrophage inflammatory protein-1α (MIP-1α), a newly discovered cytokine of chemokine family, was determined in cerebrospinal fluid (CSF) from 18 patients with multiple sclerosis (MS) and from control patients with other neurological disorders by an enzyme-linked immunosorbent assay (ELISA). The concentration of MIP-1α in CSF was significantly elevated in MS in relapse (4.4 pg/ml) compared with non-inflammatory neurological disease control samples (0.3 pg/ml) ( p < 0.0002). These concentrations in MS patients correlated well with leukocyte cell counts and protein content in CSF ( r = 0.845, p < 0.0001; r = 0.853, p < 0.0001, respectively). In other inflammatory neurological disorders such as Behçet's disease and HTLV-1 associated myelopathy, significantly increased CSF levels of MIP-1α were also observed. Chemokines are reported to play an important role in an early event of inflammation such as lymphocyte traffic. This report is the first study which confirmed the involvement of a chemokine in MS and other inflammatory neurological disorders.

Decrease in 6R-5,6,7,8-tetrahydrobiopterin content in cerebrospinal fluid of autistic patients

The levels of pterin compounds in the lumbar cerebrospinal fluid (CSF) of ten neurologically normal and twenty autistic children were measured by two different liquid chromatographic systems, a conventional and a newly developed direct method. Among pterin compounds 7,8-dihydroneopterin (NH 2) and 6 R-5,6,7,8-tetrahydrobiopterin ( R-BH 4) levels in autistic children were significantly reduced to 66.1 and 41.5%, respectively, of those found in the controls. The autistic children were divided into three subgroups according to their CSF homovanillic acid (HVA) level and age: the group with an elevated HVA level, the younger (<7 year old) group with a normal HVA level, and the older (〉7 year old) group with a normal HVA level. In comparison, the further reduction was observed in NH 2 and R-BH 4 levels in the younger group and in the 7,8-dihydropterin (PH 2) level in the group with an elevated HVA level. Thus, these results suggest that the endogenous biosynthesis of R-BH 4 in the brain may be reduced in autistic children as compared with that in neurologically normal children.

Cerebrospinal monoamine metabolites and amino acid content in patients with parkinsonian syndrome and rats lesioned with MPP+.

Monoamine metabolites and amino acid concentration in cerebrospinal fluid (CSF) of 33 untreated patients with parkinsonian syndrome, and 20 control patients without specific neurological symptoms have been compared with those obtained in cerebrospinal fluid of rats intrastriatally lesioned with 1-methyl-4-phenylpyridinium ion (MPP+) and sham operated animals. Homovanillic acid content was found to be significantly lower in patients with severe parkinsonism (motor score of UPDRS > 24), but not in patients with mild symptoms (motor score < or = 24). A correlation between the loss of striatal dopamine and the decrease in cerebrospinal homovanillic acid has been established in rats treated with MPP+. The extrapolation of these results to those obtained from human patients could be important in assessing the degree of striatal dopamine loss shown by humans with parkinsonian syndrome at the moment of clinical diagnosis. No significant differences were found between the other monoamine metabolites analyzed and free amino acid content in human and rat CSF.

The effect of a single intraperitoneal dose of hrIL-1α on substance P-, neurokinin A-, calcitonin gene-related peptide- and neuropeptide Y-like immunoreactivity in cerebrospinal fluid, plasma and knee joint synovial fluid in the rat

Substance P (SP)-, neurokinin A (NKA)-, calcitonin gene-related peptide (CGRP)- and neuropeptide Y (NPY)-like immunoreactivity (-LI) was studied in rats' cerebrospinal fluid (CSF), plasma and synovial fluid (SF) from both knee joints at 2 and 24 h following an intraperitoneal administration of 0.05 ml human recombinant interleukin-1α (i.p. hrIL-1α) or saline. Increased or decreased levels of SP-, NKA and CGRP-LI were detected in CSF and plasma, whereas NPY-LI was unaffected. In SF only CGRP-LI increased bilaterally. There was a correlation in CGRP-LI content between plasma and CSF following i.p. hrIL-1α but not between plasma and SF or CSF and SF. It can be concluded that (1) i.p. hrIL-1α activates somatosensory afferents thereby increasing SP- and CGRP-LI content in CSF and plasma and NKA-LI in CSF; 2) i.p. hrIL-1α induces a bilateral increase of CGRP-LI in SF which is not mediated through systemic circulation and is possibly a part of the general host defensive reaction.

Multiple sclerosis patients express increased levels of β-nerve growth factor in cerebrospinal fluid

We describe the measurement of β-nerve growth factor (NGF) content in cerebrospinal fluid (CSF) from multiple sclerosis (MS) patients compared with CSF from age-matched normal subjects using a specific sandwich immunoassay (ELISA). During acute attacks patients exhibit a significant increase of NGF content compared to controls. In contrast during remission the mean NGF levels in CSF markedly decrease. These results strongly indicate that increased NGF production in CSF is a characteristic feature of the MS inflammatory response.

Endogenous benzodiazepine receptor ligands in idiopathic recurring stupor

"Endozepines" are endogenous ligands for the benzodiazepine recognition sites on gamma-aminobutyric acid A receptors in the nervous system. Idiopathic recurring stupor (IRS) is a syndrome of spontaneous stupor or coma that is not associated with known metabolic, toxic, or structural abnormalities but can be reversed by flumazenil, a pure benzodiazepine antagonist. We measured endozepine-2 and endozepine-4 by high-performance liquid chromatography and radioreceptor assay in serum and cerebrospinal fluid from three patients with IRS. During episodes of stupor there was a large (up to 300-fold compared with control patients) increase of endozepine-4 content in cerebrospinal fluid and serum, but a return to normal concentrations between attacks. Endozepine-4 may contribute to, or be the cause of, IRS. The reasons for abnormal concentrations of endozepine in blood and brain are unknown.

NBQX, an AMPA antagonist, reduces glutamate-mediated brain edema

Glutamate (2.5 mg) was administered after the blood-brain barrier had been opened by a unilateral intracarotid infusion of 5 mg protamine sulfate in rats. Whereas the brain specific gravity, measured 24 h later, did not differ between the injected and non-injected side after protamine alone, glutamate significantly reduced the specific gravity in the right hemisphere indicating brain edema (P < 0.01). NBQX, a potent AMPA receptor antagonist, significantly reduced the edema (P < 0.01) and completely inhibited the glutamate mediated increase in albumin content in cerebrospinal fluid (P < 0.01).

Nonketotic hyperglycinemia: Treatment with NMDA antagonist and consideration of neuropathogenesis

Patients with neonatal-onset nonketotic hyperglycinemia have high-glycine content in cerebrospinal fluid (CSF) which is believed to be a cause of intractable neurologic manifestations. The glycine receptor was believed to be inhibitory in the central nervous system; however, a newly discovered glycine receptor is of the excitatory N-methyl- d-aspartate (NMDA) receptor type, which cannot be antagonized by strychnine. The NMDA receptor antagonist, ketamine, was administered to a patient with nonketotic hyperglycinemia; he demonstrated some improvement in hyperirritability, voluntary movement, and electroencephalographic findings. Strychnine therapy had been administered before this trial of NMDA antagonist, but without improvement. The respiratory condition improved with the reduction of the CSF glycine level after withdrawal of sodium valproate. Our findings indicate that high-glycine content in CSF may affect the brain in different ways via NMDA and classic glycine receptors.