Marked increase in cerebrospinal fluid ubiquitin in Creutzfeldt-Jakob disease.

We have established the radioimmunoassay for ubiquitin in the cerebrospinal fluid (CSF) and measured the ubiquitin concentration in CSF from 4 cases of neuropathologically verified Creutzfeldt-Jakob disease (CJD), 10 cases of multi-infarct dementia (MID), 7 cases of senile dementia of Alzheimer type (SDAT), and 18 controls. The normal values were determined to range from 7.3 to 21.0 ng/ml, 14.3 +/- 1.1 ng/ml in the mean +/- S.E.M. The CSF ubiquitin levels in the cases of MID and SDAT were 16.6 +/- 6.4 ng/ml and 21.3 +/- 6.1 ng/ml, respectively. In the cases of CJD, the CSF ubiquitin was markedly increased at the early and middle stages of the disease (230.6 ng/ml in Case 1, 107.6 ng/ml in Case 2, 212.5 ng/ml in Case 3, and 377.0 ng/ml in Case 4) and these gradually decreased as the disease progressed. The measurement of CSF ubiquitin seems useful to make an early diagnosis of CJD.     

Absence of protease-resistant prion protein in the cerebrospinal fluid of Creutzfeldt-Jakob disease

Creutzfeldt-Jakob disease (CJD), believed to be caused by a protease-resistant isoform of prion protein (PrP(Sc)), usually manifests itself as a clinically distinctive age-related dementia because of its rapid progression, occasionally accompanied by cerebellar ataxia. Recently, a variant CJD (vCJD) has been described, which has prominent early psychiatric symptoms and an earlier age of death. Although cerebrospinal fluid (CSF) is part of the extracellular fluid of the central nervous system (CNS), the bulk of its proteins are derived from the plasma and there is increasing concern about possible transmission of prion disease by blood. As investigation of CSF has played a significant role in the diagnosis and management of several neurological diseases, it was decided to characterize PrP present in the CSF of CJD individuals. Significant variation was observed in the level of PrP in the CSF from both non-CJD and CJD (including vCJD) patients, and the detected PrP forms are protease-sensitive. Using a conformation-dependent immunoassay, it was further demonstrated that the PrP detected in the CSF from CJD patients was broadly similar in conformation to that found in non-CJD patients. Taken together, the results of this study fail to demonstrate any correlation between the presence of protease-resistant PrP isoform (PrP(Sc)) in the CSF and disease manifestation.     

Increased levels of epsilon and gamma isoforms of 14-3-3 proteins in cerebrospinal fluid in patients with Creutzfeldt-Jakob disease.

We established four hybridoma cell lines producing monoclonal antibodies (MAbs) against 14-3-3 proteins. Immunoblot analysis revealed that epsilon and gamma isoforms were specifically increased in premortem cerebrospinal fluid samples from patients with sporadic Creutzfeldt-Jakob disease. Furthermore, dot immunoblot analysis showed that MAbs were more specific for native antigen than polyclonal antibodies were.     

Immunoglobulin and other proteins in the cerebrospinal fluid of patients with Alzheimer''s disease.

Immunoglobulin has been measured and studied electrophoretically in cerebrospinal fluid (CSF) from 14 patients with Alzheimer's disease and 25 undemented controls. Presence or absence of the diagnosis of Alzheimer's disease was confirmed histologically, as these were postmortem specimens. There was no increased incidence of oligoclonal IgG bands in either group, and no significant differences in the levels of IgG and albumin. Non-immunoglobulin bands were found in the gamma region in some samples from Alzheimer's disease patients and controls; such bands are not found in the CSF from younger patients. There was a significantly increased incidence of double transferrin and double tau protein bands in the Alzheimer's group, suggesting that further studies of genetic markers might be worthwhile.     

High neuron-specific enolase level of cerebrospinal fluid in the early stage of Creutzfeldt-Jakob disease

Summary The measurement of neuron-specific enolase level in serum and cerebrospinal fluid was conducted time-sequentially in an autopsy confirmed patient with Creutzfeld-Jakob disease. The level was markedly high in the early stage of the disease at which time the brain CT showed no or minimal abnormalities, while falling into the normal range in the advanced stage. This is the first report of the elevated level of neuron-specific enolase in Creutzfeldt-Jakob disease.Abbreviations CT Computed tomography - CSF Cerebrospinal fluid - CJD Creutzfeldt-Jakob disease - NSE Neuronspecific enolase - CEA Carcinoembryonic antigen - PSD Periodic synchronous discharge - PET Positron emission tomography     

Abnormal isoform of prion proteins accumulates in the synaptic structures of the central nervous system in patients with Creutzfeldt-Jakob disease.

A new method, which enabled the first immunohistochemical documentation of abnormal prion protein (PrP) in all patients with Creutzfeldt-Jakob disease (CJD), was established. This method designated as "hydrolytic autoclaving" revealed punctate PrPCJD stainings around the neuronal cell bodies and dendrites in CJD brains. These punctate stainings were almost identical with that of synaptophysin, suggesting PrPCJD accumulations in the synaptic structures. Subcellular fractionation revealed that prion protein in Creutzfeldt-Jakob disease (PrPCJD) was most concentrated in the synaptosomal fraction. In CJD patients with a long clinical course, synaptophysin immunoreactivity decreased, and synaptic PrPCJD accumulated with a wider distribution. These results suggest that synaptic PrPCJD accumulations might be responsible for the neuronal dysfunction and degeneration in CJD.     

Elevated levels of phosphorylated neurofilament proteins in cerebrospinal fluid of Alzheimer disease patients

Neurofilament (NF) subunits NF-H, NF-M and NF-L are hyperphosphorylated and elevated in Alzheimer disease (AD) brain. We investigated the level and phosphorylation states of NF subunits in lumbar cerebrospinal fluid (CSF) from living patients by bienzyme substrate-recycle enzyme-linked immunosorbent assay. We found: (i), that the levels of phosphorylated NF-H/M (pNF-H/M), non-phosphorylated NF-H/M (npNF-H/M) and NF-L were significantly higher (pNF-H/M, 12–24-fold; npNF-H/M, 3–4-fold) in neurologically healthy aged people than young control individuals; (ii), that in AD, the levels of npNF-H/M, and NF-L were similar to vascular dementia (VaD), and higher than in age-matched controls; and (iii), that the levels of pNF-H/M were significantly higher than in aged controls, non-AD neurological disorders and VaD. Based on these findings, it is suggested that the increased level of total NF proteins in CSF could be used as a marker for brain aging and neurodegenerative disorders in general, and the levels of pNF-H/M as a marker to discriminate AD from normal brain aging and as well as neurological conditions including VaD.     

抑郁症患者脑脊液异常蛋白的表达

目的:探讨抑郁症患者脑脊液蛋白组学表达的差异蛋白。方法:本研究为对照研究。研究对象是符合中国精神障碍分类与诊断标准第3版(CCMD-3)抑郁症诊断标准的8例门诊首发抑郁症患者和8例正常对照。提取研究对象的脑脊液后,应用比较蛋白组学技术分析脑脊液蛋白,所得凝胶经银染和考染后,应用ImageMaster5.0软件对图像进行分析。结果:脑脊液差异蛋白点的初步分析显示,与对照组比较,抑郁症患者脑脊液中Apo D(丰度比值0.022vs.0.321)和血浆视黄醇结合蛋白(0.032vs.0.541)2种蛋白表达下调,而Apo A-Ⅰ(0.556vs.0.020)1种蛋白表达上调。结论:抑郁症患者脑脊液上述3种蛋白的差异表达可能与抑郁症的发生有关,有可能为治疗寻找新的靶点。     

Nicotinamide-N-methyltransferase is higher in the lumbar cerebrospinal fluid of patients with Parkinson's disease

Parkinson's disease (PD) may be initiated or precipitated by endogenous toxins with a structure similar to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine in genetically-predisposed individuals. Nicotinamide N-methyltransferase (NNMT) catalyzes N-methylation of nicotinamide and other pyridines to form pyridinium ions. The protein amount of NNMT was measured in the lumbar cerebrospinal fluid of PD patients by immunoblot analysis using anti-human NNMT antibody. In younger (65 years old or younger) PD patients, the relative level of NNMT protein was significantly higher than that in younger controls. The NNMT protein was significantly affected by aging: the amount decreased along with aging in PD patients. These findings suggested that excess NNMT in the central nervous system might be implicated in the PD pathogenesis.     

Increased tau in the cerebrospinal fluid of patients with frontotemporal dementia and Alzheimer's disease

Cerebrospinal fluid (CSF) concentrations of tau protein were measured using an enzyme-linked immunosorbent assay which measures both normal and hyperphosphorylated tau. Levels of CSF tau were measured in 17 patients with Alzheimer's disease and 23 patients with frontotemporal dementia, and were compared to age-matched healthy controls. The CSF tau concentration in control subjects was 198+/-49 pg/ml and no relationship was found between age and CSF tau concentrations in these subjects. CSF tau concentrations were significantly raised in patients with both Alzheimer's disease and frontotemporal dementia when compared to healthy controls (802+/-381 pg/ml, P<0.001; 612+/-382 pg/ml, P<0.05, respectively). In neither disease did CSF tau correlate with disease severity as assessed by the Mini Mental State Examination score (MMSE). This study shows that CSF tau is significantly raised in patients with frontotemporal dementia.     

Creutzfeldt-Jakob disease prion proteins in human brains

Creutzfeldt-Jakob disease is caused by a slow infectious pathogen, or prion. We found that purified fractions from the brains of two patients with Creutzfeldt-Jakob disease contained protease-resistant proteins ranging in apparent molecular weight from 10,000 to 50,000. These proteins reacted with antibodies raised against the scrapie prion protein PrP 27-30. Rod-shaped particles were found in the brain tissue of the patients that were similar to those isolated from rodents with either scrapie or experimental Creutzfeldt-Jakob disease. After being stained with Congo red dye, the protein polymers from patients with Creutzfeldt-Jakob disease exhibited green birefringence when examined under polarized light. Our findings suggest that the amyloid plaques found in the brains of patients with Creutzfeldt-Jakob disease may be composed of paracrystalline arrays of prions similar to those in prion diseases in laboratory animals.     

Tau protein concentrations in cerebrospinal fluid of non-demented Parkinson's disease patients

We measured total tau protein concentrations in the cerebrospinal fluid (CSF) of 26 non-demented Parkinson's disease (PD) patients and 25 matched controls. When compared with controls, PD patients had similar CSF tau protein concentrations. These values were not correlated with age, age at onset of PD, duration of PD, scores of the Unified PD Rating Scale (UPDRS), and the Hoehn and Yahr staging, and were not influenced significantly by antiparkinsonian drugs. Our results suggest that CSF tau protein levels are apparently unrelated to the risk of PD.     

Proteomics of tumor-specific proteins in cerebrospinal fluid of patients with astrocytoma: Usefulness of gelsolin protein

Changes in cerebrospinal fluid (CSF) composition have been shown to accurately reflect pathological processes in the CNS, and are potential indicators of abnormal CNS states, such as tumor growth. To detect biomarkers in high-grade astrocytomas, the differential expression of proteins in the cerebrospinal fluid was analyzed from two cases each of diffuse astrocytoma (grade II), and glioblastoma (grade IV) using agarose 2-D gel electrophoresis (2-DE). It was found that the expression of gelsolin protein decreased with histological grade. To examine whether gelsolin is a useful indicator of tumor aggressiveness or patient outcome, its expression was further studied on immunohistochemistry in 41 formalin-fixed and paraffin-embedded astrocytomas. The positive cell rate of gelsolin in tumors was 59.4% in grade II, 30.0% in grade III and 29.4% in grade IV, respectively. Gelsolin expression was significantly lower in high-grade astrocytomas (grade III or IV) than in low-grade astrocytomas (grade II; P < 0.05). Moreover, in astrocytomas the overall survival of patients in the low-expression group was significantly poorer than in the high expression group ( P < 0.05). These data suggest that gelsolin is a prognostic factor in astrocytoma.     

Total antioxidant capacity of cerebrospinal fluid is decreased in patients with motor neuron disease

Oxidative stress has been associated with motor neuron disease (MND). The human body has several antioxidant defense systems to repair the damage caused by oxidative stress. The activity of these systems is thought to be reduced in neurodegenerative diseases, which may increase the level of oxidative damage and be a contributing factor to motor neuron death. In the present study, we compared the total antioxidant capacity (TAC) of human serum and cerebrospinal fluid (CSF) of MND patients with that of a control group including patients with migraine, tension headache and psychiatric disorders. Within-subject serum and CSF TAC were strongly correlated (r=0.639; p=0.000), and CSF TAC was significantly lower in MND patients as compared to controls after adjustment for known influencing factors (112.7 micromol Fe/L+/-11.7 versus 135.2 micromol Fe/L+/-19.7; p=0.012). No differences in serum or CSF TAC were observed among the clinical forms of MND considered in this work. In conclusion, the CSF TAC was strongly correlated with serum TAC, and a decrease in CSF TAC was demonstrated in MND patients compared to controls that was not independent from serum antioxidants, this translating in a systemic (but prevailing in the CNS) oxidative damage in this pathology.     

Increase of neuron-specific enolase in patients with Creutzfeldt-Jakob disease

Creutzfeldt-Jakob disease (CJD) is a rare neurodegenerative human disorder with an incidence of one case per 1000000 per year. Recently new diagnostic tests such as neuron-specific enolase (NSE), S-100, tau-protein and protein 14-3-3 have been established as markers in prion diseases. NSE is elevated in case of rapid nerve cell loss so quantitative measurement of NSE in cerebrospinal fluid (CSF) might correlate with the disease progression. To further evaluate this hypothesis we analysed longitudinal CSF samples from 16 CJD patients. The first spinal tap was taken two weeks after the first clinical signs of a neurodegenerative disorder. This showed an elevation of NSE which continued during the course of the disease. Longitudinal examination of neuron-specific enolase in cerebrospinal fluid therefore may be useful for differentiation between CJD and other dementias.     

Molecular size distribution of somatostatin-like immunoreactivity in the cerebrospinal fluid of patients with degenerative brain disease

The molecular size distribution of somatostatin-like immunoreactivity (SLI) in the cerebrospinal fluid (CSF) of patients with brain disease was investigated by separation with a Sephadex G-25 superfine column and subsequent radioimmunoassay of the eluate. Marked heterogeneity of SLI in the CSF of control subjects as well as in demented patients, was observed. Controls and schizophrenics exhibited an SLI distribution pattern consisting mainly of two pronounced peaks: the first eluting with the void volume of the column; the second being compatible with a peptide of N-terminally extended somatostatin-14. SLI from the CSF of patients with senile dementia of the Alzheimer type (SDAT), multi-infarct dementia (MID) and normal pressure hydrocephalus (NPH) showed the same two peaks found in controls and schizophrenics; and in addition, a third peak co-eluting with somatostatin-14. However, this peak was more pronounced in patients with SDAT and MID than in patients with NPH. Re-chromatography of G-25 sf void volume immunoreactivity afforded two fractions of an apparent molecular weight of about 10,000 daltons and 15,500 daltons, respectively.     

Alteration of 8-hydroxyguanosine concentrations in the cerebrospinal fluid and serum from patients with Parkinson's disease

In order to investigate the possible role of oxidative RNA damage in the pathogenesis of Parkinson's disease (PD), the concentrations of the oxidative stress marker 8-hydroxyguanosine (8-OHG) were measured in the cerebrospinal fluid (CSF) and the serum of patients with PD and control subjects. The concentration of 8-OHG in CSF in PD patients was approximately three-fold that in controls (P < 0.001). The concentration of 8-OHG in CSF decreased significantly with the duration of disease (r(s) = -0.46, P < 0.05). However, the concentration of 8-OHG in serum was not significantly altered in PD patients compared to that in controls. In addition, the concentration of 8-OHG in CSF showed no correlation with that in serum in both the controls and PD patients suggesting that the 8-OHG concentrations in the CSF do not reflect those in serum and may be probably reflect those in brain tissue. These in vivo findings suggest a possible role of 8-OHG and increased oxidative RNA damage in the early stage of the development of PD.