Prostaglandin E2 is increased in amyotrophic lateral sclerosis patients

OBJECTIVES: Oxidative stress and glutamate-mediated excitotoxicity may play an important role in the etiopathogenesis of amyotrophic lateral sclerosis (ALS). Prostaglandin E2 (PGE2) activity can be associated with motor neuron death by inducing free radical formation and glutamate release from astrocytes. The aim of this study was to determine PGE2 concentration in the serum and cerebrospinal fluid (CSF) of ALS patients. MATERIAL AND METHODS: PGE2 concentration was measured by the enzyme-linked immunosorbent method in the serum and CSF from ALS and control group patients. RESULTS: Serum and CSF PGE2 concentration was significantly higher in the whole group of ALS patients compared with the control group patients (P < 0.05). There was no relationship between PGE2 concentration and clinical parameters of the disease, such as clinical state, type of ALS onset, and duration of the disease (P > 0.05). A significant correlation between CSF PGE2 concentration and age of control group patients was found (P < 0.05). CONCLUSIONS: A significant increase in serum and CSF PGE2 concentration, in ALS patients observed in this study, indicates that PGE2 may play a role in neurodegeneration of ALS through oxidative damage of neurons and glutamate-mediated excitotoxicity. It suggests that inhibition of PGE2 synthesis could prevent motor neuron death. However, serum and CSF PGE2 cannot be a marker of the type of ALS onset, clinical state of patients, or the duration of the disease.     

Endogenous protectant kynurenic acid in amyotrophic lateral sclerosis

OBJECTIVES: Excitotoxicity may play a role in neurodegeneration in amyotrophic lateral sclerosis (ALS). Kynurenic acid (KYNA), an endogenous antagonist of excitatory amino acid receptors, may inhibit excitotoxic lesions. The aim of this study was to determine the concentration of KYNA in ALS patients. MATERIAL AND METHODS: KYNA was measured by high-performance liquid chromatography in the serum and cerebrospinal fluid (CSF) from ALS and control patients. RESULTS: Our study revealed that CSF KYNA concentration was significantly higher in patients with bulbar onset of ALS compared to controls, and compared to patients with limb onset of the disease. CSF KYNA was also higher in patients with severe clinical status compared to controls. Serum KYNA was significantly lower in ALS patients with severe clinical status compared to controls, and compared to patients with mild clinical status. There were no significant differences in CSF and serum KYNA concentration between the whole ALS group of patients and controls. There was no difference in CSF KYNA concentration between males and females, and there was no correlation between KYNA concentration and age of patients, and duration of ALS. CONCLUSIONS: An increased CSF KYNA concentration in patients with bulbar onset of ALS and in patients with severe clinical status may indicate neuroprotective role of KYNA against excitotoxicity. The difference of KYNA concentration in CSF of patients with bulbar and limb onset of ALS suggests that these two variants of motor neuron disease may have different etiopathogenetic mechanisms.     

Cerebrospinal fluid Flt3 ligand level in patients with amyotrophic lateral sclerosis

OBJECTIVES: The Flt3 ligand (FL) is a cytokine with a neurotrophic and antiapoptotic activity in the central nervous system that induces the survival of neurons. The aim of the study was to measure levels of FL in amyotrophic lateral sclerosis (ALS) patients. MATERIALS AND METHODS: The study involved 23 ALS patients and 23 people in the control group. The measurement of FL in the cerebrospinal fluid (CSF) and serum was performed by the enzyme-linked immunosorbent method. RESULTS: Results showed that CSF FL levels were significantly increased in ALS patients compared with the controls (P < 0.05) but the serum levels of this cytokine did not differ from the controls (P > 0.05). There was no significant correlation between CSF and serum FL levels and clinical parameters of ALS (P > 0.05). The difference in CSF/serum ratio of FL between ALS patients and controls was not statistically significant (P > 0.05). CONCLUSION: An increase in CSF FL levels in ALS patients, observed in this study, could be a compensative response for neurodegeneration but may also reflect increased diffusion of this cytokine into the central nervous system caused by blood-CSF barrier dysfunction.     

Modulators of cystein proteases and cell-death markers in the cerebrospinal fluid of patients with amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease characterized by selective damage of motor neurons in the brain and spinal cord. The aim of the current study was to reveal new specific markers of neurodegeneration in the cerebrospinal fluid (CSF) of ALS patients. We measured the activities of substances that modulate the activity of cysteine proteases (calpain, caspase, and cathepsin B) in the CSF of ALS patients. The CSF of ALS patients, in contrast to the CSF of patients with multiple sclerosis, inhibits calpain and cathepsin B significantly more strongly, as compared to the control CSF. The LDH activity and the concentration of neuron-specific enolase (NSE) in the CSF of control patients and ALS patients did not differ. The concentration of neurofilament heavy chains in the CSF was significantly higher in the ALS patients as compared to the control group. The albumin index increased in 50% of all ALS patients. We also have shown a correlation between the cathepsin inhibitor activity, the concentration of neurofilament heavy chains, and the albumin index and the clinical findings of the disease. According to our data, the concentration of neurofilament heavy chains is a sensitive marker of neurodegenerative process during ALS. An increase in the CSF inhibiting activity with respect to cathepsin B and calpain is specific for ALS and may confirm the role of an imbalance of proteolytic systems in the pathogenesis of this disease.     

CSF neurofilament protein analysis in the differential diagnosis of ALS

Background

Cerebrospinal fluid (CSF) biomarkers have been studied to differentiate between patients with ALS and neurological controls, but not in comparison to clinically more relevant disorders mimicking ALS.

Methods

In this retrospective study, CSF concentrations of various brain-specific proteins were analyzed in patients with ALS (n = 32) and ALS-mimic disorders (n = 26).

Results

CSF concentrations of neurofilament light (NFL) and heavy chain (NFHp35), but not other brain-specific proteins, were significantly higher in patients with ALS than in patients with an ALS-mimic disorder, however with maximum sensitivity or specificity of 80 %. The mean CSF level of NFHp35 was 781 ng/L in the ALS group vs. 338 ng/L in the ALSmimic disorders group and for NFL the mean CSF levels were 62 ng/L vs. 24 ng/L.

Conclusion

Although CSF concentrations of NFL and NFHp35 are higher in patients with ALS, the diagnostic accuracy for differentiating ALS from ALS-mimic disorders seems insufficient. Our results suggest that, in the clinical work-up of patients suspected of ALS, application of CSF analysis alone is limited but may have potential in combination with other clinical and electrophysiological markers.     

Elevation of MCP-1 and MCP-1/VEGF ratio in cerebrospinal fluid of amyotrophic lateral sclerosis patients

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease with progressive cell death of upper and lower motor neurons. In this study, we measured monocyte chemotactic protein-1 (MCP-1) and vascular endothelial growth factor (VEGF) levels in cerebrospinal fluid (CSF) and serum by enzyme-linked immunosorbent assay (ELISA) in 42 ALS patients, and compared these levels with those of control subjects with other neurodegenerative disorders or with those of normal controls. MCP-1 levels in CSF were significantly higher in ALS patients than in the control group. VEGF levels in CSF tended to be lower in ALS patients than in the control group, but not significantly. A positive correlation was found between MCP-1 levels in CSF of ALS patients and the total Norris scale. The elevation of MCP-1/VEGF ratio in CSF was more specific to ALS patients compared to other neurological diseases such as Parkinson's disease (PD) and spinocerebellar ataxia (SCA) and to controls. Our data suggested that both MCP-1 levels and MCP-1/VEGF ratio in CSF may be useful markers for the clinical diagnosis of ALS.     

Cerebrospinal fluid vascular endothelial growth factor in patients with amyotrophic lateral sclerosis

Oxidative stress and glutamate-mediated toxicity may play an important role in the etiopathogenesis of amyotrophic lateral sclerosis (ALS). The vascular endothelial growth factor (VEGF) is a neuroprotective cytokine activated by hypoxia. The aim of this study was to measure VEGF levels in the cerebrospinal fluid (CSF) of ALS patients. The study concerned 30 ALS patients and 30 control subjects. The VEGF was measured by the enzyme-linked immunosorbent assay. The results have shown that CSF VEGF levels are significantly increased in patients with long duration of ALS and in patients with limb-onset of the disease compared with controls (P<0.05). Moreover, the type of ALS patients’ subgroup significantly influences CSF VEGF levels (P=0.05). The CSF VEGF levels were significantly increased in patients with limb-onset compared to patients with bulbar-onset of ALS, and in patients with long duration of ALS compared to patients with its short duration (P<0.05). There was a significant correlation between CSF VEGF levels and duration of ALS (P<0.05). It seems that a significant increase in CSF VEGF levels in patients with limb-onset of ALS and in patients with long duration of the disease may have a protective role against glutamate-mediated toxicity and oxidative damage of motor neurons. However, the conclusions are limited due to relatively small subgroups of ALS patients and by lack of a control group consisting of healthy persons. Further investigations could help to confirm the results from this preliminary report.     

Elevation of MCP-1 and MCP-1/VEGF ratio in cerebrospinal fluid of amyotrophic lateral sclerosis patients

Abstract

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease with progressive cell death of upper and lower motor neurons. In this study, we measured monocyte chemotactic protein-1 (MCP-1) and vascular endothelial growth factor (VEGF) levels in cerebrospinal fluid (CSF) and serum by enzyme-linked immunosorbent assay (ELISA) in 42 ALS patients, and compared these levels with those of control subjects with other neurodegenerative disorders or with those of normal controls. MCP-1 levels in CSF were significantly higher in ALS patients than in the control group. VEGF levels in CSF tended to be lower in ALS patients than in the control group, but not significantly. A positive correlation was found between MCP-1 levels in CSF of ALS patients and the total Norris scale. The elevation of MCP-1/VEGF ratio in CSF was more specific to ALS patients compared to other neurological diseases such as Parkinson's disease (PD) and spinocerebellar ataxia (SCA) and to controls. Our data suggested that both MCP-1 levels and MCP-1/VEGF ratio in CSF may be useful markers for the clinical diagnosis of ALS.     

CSF concentrations of adipsin and adiponectin in patients with amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis (ALS) is described as a neurodegenerative disorder. However, neuroinflammation and chemokine expression are prominent pathological finding at sites of injury. Adipsin and adiponectin are molecules that are implicated in the pathogenesis of neurodegenerative and neuroimmune disorders. Adipsin and adiponectin concentrations were determined in the CSF of ALS patients and controls and the relationship of these chemokines with clinical severity and disease duration in ALS was determined. Seventy-seven ALS patients (mean age 49.5 ± 10.4 years) (mean body mass index 23.5 ± 4.5) were included. Twenty patients had bulbar, 53 spinal, and four bulbospinal onset ALS. Median adipsin CSF level was 12,650.94 pg/ml in ALS patients and 3290.98 pg/ml in controls (p < 0.001). Median adiponectin CSF level was 4608 pg/ml in ALS patients and 3453 pg/ml in controls (p = 0.1). No differences were observed in disease duration, progression rate or disease severity. There was a significant positive correlation between adipsin and adiponectin concentrations (r = 0.379, p = 0.01). No correlation with age, body mass index or ALFRS-R score was found. Adipsin was significantly elevated in CSF, suggesting that this chemokine might have a role in ALS pathogenesis. Adiponectin showed a trend towards higher concentrations, but failed to reach statistical significance. Due to the clinical heterogeneity in our cohort, these chemokines do not appear to be associated with disease duration or severity.     

CSF neurofilament proteins as diagnostic and prognostic biomarkers for amyotrophic lateral sclerosis

Elevated cerebrospinal fluid (CSF), Neurofilament Light (NF-L) and phosphorylated Heavy (pNF-H) chain levels have been found in Amyotrophic Lateral Sclerosis (ALS), with studies reporting a correlation of both neurofilaments (NFs) with the disease progression. Here, we measured NF-L and pNF-H concentrations in the CSF of ALS patients from a single tertiary Center and investigated their relationship with disease-related variables. A total of 190 ALS patients (Bulbar, 29.9%; Spinal, 70.1%; M/F = 1.53) and 130 controls with mixed neurological diseases were recruited. Demographic and clinical variables were recorded, and ΔFS was used to rate the disease progression. Controls were divided into two cohorts: (1) patients with non-inflammatory neurological diseases (CTL-1); (2) patients with acute/subacute inflammatory diseases and tumors, expected to lead to significant axonal and tissue damage (CTL-2). For each patient and control, CSF was taken at the time of the diagnostic work-up and stored following the published guidelines. CSF NF-L and pNF-H were assayed with commercially available ELISA-based methods. Standard curves (from independent ELISA kits) were highly reproducible for both NFs, with a coefficient of variation < 20%. We found that CSF NF-L and pNF-H levels in ALS were significantly increased when compared to CTL-1 (NF-L: ALS, 4.7 ng/ml vs CTL-1, 0.61 ng/ml, p < 0.001; pNF-H: ALS, 1.7 ng/ml vs CTL-1, 0.03 ng/ml, p < 0.0001), but not to CTL-2. Analysis of different clinical and prognostic variables disclosed meaningful correlations with both NF-L and pNF-H levels. Our results, from a relatively large ALS cohort, confirm that CSF NF-L and pNF-H represent valuable diagnostic and prognostic biomarkers in ALS.     

Reduced angiotensin II levels in the cerebrospinal fluid of patients with amyotrophic lateral sclerosis

Background – Recent studies suggest that angiotensin II, a major substrate in the renin–angiotensin system, protects neurons through stimulation of its type 2 receptors. However, quite a few clinical studies of angiotensin II levels have shown their relation to disease severity in neurodegenerative disorders, including amyotrophic lateral sclerosis (ALS). Aims of the study – To clarify the significance of angiotensin II in ALS. Methods – We assayed angiotensin II concentrations in cerebrospinal fluid (CSF) samples from 23 patients with ALS, nine patients with spinocerebellar degeneration (SCD) and 24 control individuals. We evaluated the disability levels of patients with ALS using the Revised ALS Functional Rating Scale (ALSFRS‐R) and calculated the disease progression rate (DPR). Results – CSF angiotensin II levels were significantly lower in the ALS group compared with that in the control group (P = 0.00864), and showed a significant positive correlation with scores on the ALSFRS‐R, and a significant negative correlation with the DPR. Conclusions – In the present study, we reveal for the first time that angiotensin II levels in the CSF from patients with ALS are significantly reduced and significantly associated with disease severity and progression rate. These findings suggest that reduced levels of intrathecal angiotensin II may play a role in ALS.     

Serum caspase-9 levels are increased in patients with amyotrophic lateral sclerosis

It is known that apoptosis may play a role in the pathophysiology of amyotrophic lateral sclerosis (ALS). Moreover, caspase-9 is implicated in the apoptosis pathway. The aim of the study was to investigate caspase-9 levels in serum of patients with ALS. The study involved 30 patients with ALS and 30 patients from the control group. The serum caspase-9 levels were measured using the enzyme-linked immunosorbent method. The study showed that caspase-9 levels are significantly increased in serum of the patients with ALS comparing to the control group (p < 0.05). There was a significant correlation of serum caspase-9 levels with severity of clinical state of ALS patients and duration of the disease (p < 0.05). The results indicate that caspase-9 may be implicated in pathomechanism of neurodegeneration in ALS.     

肌萎缩侧索硬化患者的脑脊液蛋白及髓鞘碱性蛋白相关研究

目的探讨肌萎缩侧索硬化(ALS)患者的脑脊液蛋白、髓鞘碱性蛋白(MBP)水平及脑脊液蛋白与临床特征之间的关系。方法回顾性研究行腰穿查脑脊液的29例确诊ALS患者,检测其脑脊液蛋白及MBP水平,并按性别、年龄、病程、起病部位及临床功能评分〔肌萎缩侧索硬化功能分级量表(ALS-FRS)评分〕等不同临床特征分组,分析不同临床特征对脑脊液蛋白水平的影响。结果 29例ALS患者脑脊液蛋白水平为(0.43±0.15)g/L,其中脑脊液蛋白轻度增高患者9例(31%),最高为0.89g/L;不同性别〔男(0.42±0.15)g/L,女(0.45±0.18)g/L,t=0.501,P=0.620〕、年龄〔60岁组(0.43±0.17)g/L,≥60岁(0.44±0.13)g/L,t=0.141,P=0.889〕、病程〔1年组(0.37±0.11)g/L,≥1年(0.49±0.17)g/L,t=-2.23,P=0.054〕、起病部位〔球部起病组(0.38±0.11),肢体起病组(0.45±0.17),t=0.330,P=0.743〕、ALS-FRS评分〔30分组(0.42±0.16)g/L,≤30分组(0.44±0.16)g/L,t=0.092,P=0.928〕分组间比较,脑脊液蛋白水平差异均无统计学意义。29例患者中13例进行了脑脊液MBP检测,MBP水平(1.66±0.78)nmol/L,13例患者MBP水平均增高,最高达3.39nmol/L。MBP水平与脑脊液蛋白水平无相关性(R=0.198,P=0.517)。结论 ALS患者脑脊液蛋白增高多见。部分ALS患者脑脊液MBP水平增高,但与脑脊液蛋白水平无相关性。     

Phosphorylated neurofilament heavy subunits as a marker of neurodegeneration in demyelinating diseases of the CNS

Neurofilaments are used as biomarkers of neurodegeneration, i.e., neuroinflammatory destruction of neurons, in various diseases of the nervous system. The purpose of our study is to clarify data on the pathogenesis and clinical prognosis of chronic and rare monophase demyelinating diseases. The content of phosphorylated neurofilament heavy subunits (pNFH) was measured in the cerebrospinal fluid (CSF) and blood serum using ELISA. The following groups of patients were studied: remittent multiple sclerosis (RMS, n = 56), secondary progressive multiple sclerosis (SPMS, n = 20), acute disseminated encephalomyelitis (ADEM, n = 7), neuromyelitis optica (NMO, n = 5), amyotrophic lateral sclerosis (ALS, n = 26), and the control group (n = 26). We found significant differences in the content of pNFH in the CSF between the RMS, ADEM, ALS groups and the control group; however, we did not observe any differences between the RMS, ADEM, and ALS groups per se. We did not find any differences in the contents of pNFH in the blood serum or any correlations of the pNFH levels with clinical features of diseases in these groups of patients. Based on this index, we conclude that neurodegeneration occurs in RMS, ADEM, and ALS. The pNFH content should be studied in larger cohorts of patients and at strictly defined time points.     

T4, T3 and rT3 levels in serum and cerebrospinal fluid of patients with amyotrophic lateral sclerosis

Summary Thyronine (T₄), triiodothyronine (T₃), and reversetriiodothyronine (rT₃) levels were evaluated in cerebrospinal fluid (CSF) and in serum of 12 patients with definite amyotrophic lateral sclerosis (ALS) by specific radioimmunoassays. Circulating microsomal and thyroglobulin antibodies were also evaluated. In all patients serum levels of T₄, T₃ and rT₃ were within normal limits. In CSF, the rT₃ levels were significantly elevated to 0.118 g/l (mean), the T₄ levels were not significantly elevated, and the T₃ levels were below the detection limit of 0.03 g/l. A correlation between the elevated rT₃ levels in CSF and the severity or type of ALS could not be demonstrated by this study. The antithyroid antibodies (thyroglobulin antibodies, microsomal antibodies) showed normal titres and did not suggest disturbances of thyroid autoimmunity in the patients with ALS.     

Increased cerebrospinal fluid cAMP levels in Alzheimer's disease

Since increasing evidence suggests that upregulation of the cAMP-second messenger system may be implicated in Alzheimer's disease neurodegeneration, we have compared the cAMP and cGMP levels in cerebrospinal fluid (CSF) from patients with dementia of the Alzheimer type (DAT, n=10) with those from nondemented age-matched controls (n=10). Our results show that cAMP levels, but not cGMP, are significantly (p<0.01) elevated in CSF from patients with DAT compared to those from nondemented controls. Moreover, a linear regression analysis demonstrated a significant correlation (r=0.62; p<0.01) between cAMP and tau protein levels in CSF when controls and patients with DAT were studied together. These results suggest that upregulation of cAMP-signaling pathway is implicated in Alzheimer's disease physiopathology.     

Increased TDP-43 protein in cerebrospinal fluid of patients with amyotrophic lateral sclerosis

There is mounting pathological, biochemical and genetic evidence that the metabolism and aggregation of the 43-kDa transactive response (TAR)-DNA-binding protein (TDP-43) play a crucial role in the pathogenesis of sporadic and some forms of familial amyotrophic lateral sclerosis (ALS). Recently, it was reported using an ELISA system that elevated levels of TDP-43 were detected in plasma samples from patients with Alzheimer’s disease and frontotemporal dementia, compared to healthy controls. To determine whether quantification of TDP-43 in cerebrospinal fluid (CSF) is potentially informative in the diagnosis of ALS, we measured the concentration, by a similar ELISA method, of TDP-43 in CSF from 30 patients with ALS (diagnosed according to the revised El Escorial criteria) and 29 age-matched control patients without any neurodegenerative disease. We found that, as a group, the ALS patients had significantly higher levels of TDP-43 in their CSF than the age-matched controls (6.92 ± 3.71 ng/ml in ALS versus 5.31 ± 0.94 ng/ml in controls, p < 0.05), with levels of TDP-43 in CSF elevated beyond 95% upper confidence level for the control group in six (20%) of the patients with sporadic ALS. All the six patients with higher levels of CSF TDP-43 were examined within 10 months of the onset of illness. The patients examined within 10 months of onset showed significantly higher levels of CSF TDP-43 (8.24 ± 4.72 ng/ml) than those examined after 11 months or more of onset (5.41 ± 0.66 ng/ml, p < 0.05). These results suggest that the levels of TDP-43 in CSF may increase in the early stage of ALS. We also confirmed the existence of the TDP-43 protein in CSF from some patients with ALS, and a control subject, by western blotting of proteins immunocaptured from the CSF samples. Raised TDP-43 levels in the CSF may preempt the formation of TDP-43 pathology in the central nervous system, or correlate with early-stage TDP-43 pathology, and accordingly be a biomarker for the early stage of ALS.     

Low concentrations of glutamate induce apoptosis in cultured neurons: implications for amyotrophic lateral sclerosis

Evidence is accumulating that excessive glutamate concentration in the extracellular space is neurotoxic and plays a role in amyotrophic lateral sclerosis (ALS). However, the published results on glutamate levels in cerebrospinal fluid (CSF) and on glutamate-mediated toxicity of CSF in ALS disease remain controversial. In this report, we studied CSF from patients with sporadic ALS and controls to determine glutamate concentrations, and then analyzed the neurotoxic effect of glutamate at the concentrations present in CSF from ALS patients on cultured cortical neuronal cells. Our study shows that glutamate, at the concentrations found in CSF from ALS patients (5.8 microM), diminished cell viability and increased apoptosis determined by the fluorescent DNA-binding dye Hoechst 33342 as well as by Terminal deoxynucleotidyl transferase (TdT)-mediated dUTP Nick End-Labeling (TUNEL) reaction in cultured neuronal cells. However, glutamate concentrations as those found in CSF from controls (2.8 microM or below) did not induce any effect. Both significant glutamate-induced effects were inhibited in the presence of NBQX (2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(f)quinoxaline-2,3-dione), an alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA)/kainate-sensitive glutamate receptor antagonist. These results demonstrate that AMPA/kainate receptors are involved in the glutamate-mediated neurotoxic effects on cultured neurons, according to reports that implicate these receptors in ALS disease. We conclude that the glutamate-mediated neuronal apoptosis through AMPA/kainate receptors could occur in ALS patients who have elevated CSF glutamate concentration.