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.     

Lead concentrations in blood, plasma, erythrocytes, and cerebrospinal fluid in amyotrophic lateral sclerosis

The purpose of the investigation was to elucidate the repeatedly discussed relationship between chronic lead intoxication and ALS. The following mean lead concentrations were determined in 9 patients with ALS: 8.65 +/- 3,91 micrograms/100 ml in the blood, 0.97 +/- 0.78 microgram/100 ml in the plasma, 19.15 +/- 5.0 micrograms/100 ml in the erythrocytes, and 0.89 +/- 0.44 microgram/100 ml in the cerebrospinal fluid. These values did not differ appreciably from the controls with 7.91 +/- 3.83 micrograms/100 ml (n = 14) in the blood, 1.13 +/- 0.46 microgram/100 ml (n = 10) in the plasma, 18.96 +/- 12.63 micrograms/100 ml (n = 10) in the erythrocytes, and 0.85 +/- 0.91 microgram/100 ml (n = 15) in the cerebrospinal fluid. These findings do not support the assumption of lead poisoning as a pathogenetic factor in ALS.     

散发性肌萎缩侧索硬化病人脑脊液中3-硝基酪氨酸水平增高

目的 探讨自由基氧化损伤在散发性肌萎缩侧索硬化的发病机制中有无作用. 方法 散发性肌萎缩侧索硬化病人15例,神经系统正常的外科手术腰麻病人20例,采集脑脊液后用高效液相色谱法进行检测分析.比较散发性肌萎缩侧索硬化病人和神经系统正常的受试者脑脊液中3-硝基酪氨酸、酪氨酸含量及3-硝基酪氨酸与酪氨酸比值.结果 散发性肌萎缩侧索硬化病人脑脊液中3-硝基酪氨酸含量及3-硝基酪氨酸与酪氨酸比值与对照组相比增高有显著性差异病人组与对照组脑脊液中3-硝基酪氨酸含量分别为(228.52±124.30) nmol/L和(112.86±47.10) nmol/L,P<0.01;两组的3-硝基酪氨酸与酪氨酸比值分别为(7.60±4.38)×10-3和(3.19±1.20) ×10-3,P<0.01.结论 由过硝酸根介导的自由基氧化损伤在散发性肌萎缩侧索硬化的发病机制中具有重要作用.     

Epidermal growth factor in human cerebrospinal fluid: reduced levels in amyotrophic lateral sclerosis

Summary Epidermal growth factor (EGF), a mitogenic peptide, is widely distributed within the brain and endocrine cells of the gastro-intestinal tract. Using EGF radioreceptor assay, the EGF level was measured in lumbar cerebrospinal fluid from five patients with amyotrophic lateral scerlosis (ALS) and seven patients with intervertebral disc disease as a control group. The patients with ALS showed reduced EGF levels to 662.4±207 pg/ml as compared with controls 1013±182.8 pg/ml (P<0.02). These results indicate a possible EGF involvement in the pathogenesis of ALS.This work is partially supported by M.Z. V/31     

Intrathecal thyrotropin-releasing hormone therapy of amyotrophic lateral sclerosis

Summary Six patients with amyotrophic lateral sclerosis were given from 800 to 4000 g of thyrotropin-releasing hormone (TRH) intrathecally for a period of 2–6 months. The progressive course of this disease, manifested by increasing atrophy, paralysis and disability score, was not altered. This supports the hypothesis that the decrease in TRH content in the anterior horn region is secondary to the cellular destruction. TRH appears to play no significant role in the pathogenesis of amyotrophic lateral sclerosis     

Cerebrospinal fluid levels of alpha-tocopherol in amyotrophic lateral sclerosis

Summary. We compared CSF and serum levels, and the CSF/serum ratio of alpha-tocopherol (vitamin E), measured by HPLC, in 30 patients with sporadic amyotrophic lateral sclerosis (SALS) and 78 matched controls. The mean CSF and serum vitamin E levels did not differ significantly between the 2 study groups. These values were not influenced by the clinical form (spinal versus bulbar) of SALS. CSF alpha-tocopherol levels did not correlate with age, age at onset, and duration of the disease. These results suggest that CSF and serum alpha-tocopherol concentrations are unrelated with the risk for ALS. Received February 25, 1998; accepted April 23, 1998     

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.     

Neurofilament phosphorylation is enhanced in cultured chick spinal cord neurons exposed to cerebrospinal fluid from amyotrophic lateral sclerosis patients

Amyotrophic lateral sclerosis (ALS), a neurodegenerative disease, is characterized by degeneration of lower and upper motor neurons. Serum and cerebrospinal fluid (CSF) of ALS patients have been found to exert toxic effects on neurons in culture. We report here increased phosphorylation of neurofilaments (NF) in the soma of chick spinal cord neurons in culture when exposed to CSF of ALS patients. Spinal cord neurons were cultured from 10-day embryonic chick and exposed to culture medium supplemented with CSF or serum (10%) from ALS and non-ALS patients for 48 h. There was a significant increase in the number of neuronal soma staining with antibodies against phosphorylated NF, following exposure to CSF from ALS patients. Such an increase, however, was not observed in cultures exposed to serum from ALS patients and also serum and CSF from non-ALS patients. These results suggest that the CSF of ALS patients may contain factor(s) which induces aberrant phosphorylation of NF in the soma, a probable forerunner to the formation of neurofibrillary tangles and eventual degeneration of neurons.Supported by a research grant from the National Institute of Mental Health and Neuro Sciences, Bangalore, India     

Cerebrospinal fluid from amyotrophic lateral sclerosis has no effect on intracellular free calcium in cultured cortical neurons

The data from the literature regarding the presence of a neurotoxic factor in amyotrophic lateral sclerosis (ALS) plasma or cerebrospinal fluid (CSF) remain controversial. As a new approach to this question, we have studied the effect of CSF from ALS patients on the temporal dynamics of the intracellular free calcium concentration ([Ca²⁺]_i) of murine cortical neurons in cultures using Fura-2 fluorescence videomicroscopy and single-cell imaging. CSF from seven ALS patients and controls was added at dilutions up to 20% to cortical neuronal cultures. The in vitro inhibition of CSF on [³H]kainic acid binding showed that the CSF did not contain any substances other than glutamate itself in larger amounts. At the concentrations used, the CSF did not have any effect on [Ca²⁺]_i or on the neuronal responsiveness as defined by the ability of the cells to respond with a transient increase in [Ca²⁺]_i to depolarization induced by KCl. The disturbance of the intracellular calcium homeostasis is one of the key mechanisms of action of excitotoxic compounds mediating delayed neuronal cell death by stimulation of glutamate receptor subtypes. In this study, CSF from ALS patients did not induce immediate rises in [Ca²⁺]_i or disturbances of the intracellular calcium homeostasis when measured over a period of 2 h.     

Cerebrospinal fluid filtration in amyotrophic lateral sclerosis

By means of a randomized, controlled and open study the authors wanted to find out if cerebrospinal (CSF)-filtration was of substantial benefit to patients with sporadic amyotrophic lateral sclerosis (SALS). Five SALS patients, aged 51-75 years, being treated with riluzole underwent CSF-filtration daily over five days (group A). Five other SALS patients, aged 52-70 years, were treated only with riluzole (group B). Although all five patients in the first group reported a subjective benefit following CSF- filtration, the Norris score, the Frenchay score, the vital capacity, the ulnar nerve F-wave persistence and the peak-ratio of the brachial biceps and anterior tibial muscles did not change significantly after five days of therapy, either in group A or in group B. In conclusion, filtration of 200-250 ml CSF daily, over five days, does not seem to have a substantial therapeutic effect in patients with SALS. Copyright Lippincott Williams & Wilkins     

Respiratory function in amyotrophic lateral sclerosis

Abstract. The aim of this study was to examine the vital capacity (FVC) and forced expiratory volume in 1 s (FEV 1) in relation to the site of amyotrophic lateral sclerosis (ALS) onset and the duration of the disease. Respiratory involvement is the principal cause of death in ALS patients. The study was conducted at the Department of Neurology, University School of Medicine in Lublin. The study comprised 18 ALS patients. The average duration of ALS was 12 months. The patients were divided into two groups according to the site of ALS onset and into two groups according to the duration of the disease. FVC was significantly higher in the group of patients with a limb onset than in the group of patients with a bulbar onset of the disease. The study has shown respiratory function disturbances in ALS patients. FVC significantly depends on the site of ALS onset but not on the duration of the disease.     

Decreased cerebrospinal fluid cGMP levels in patients with amyotrophic lateral sclerosis

Summary. The role of cyclic guanosine 5 monophosphate (cGMP) in neurodegeneration of motor neurons in amyotrophic lateral sclerosis (ALS) is still controversial. The aim of this study was to measure levels of cGMP in cerebrospinal fluid (CSF) of patients with ALS, and to investigate whether there is a relationship between CSF cGMP levels and clinical parameters of the disease. The study involved 30 ALS and 20 control group patients. The CSF cGMP was measured by the enzyme-linked immunosorbent assay. The results showed that levels of CSF cGMP were significantly decreased in the group of ALS patients compared to controls and did not depend on clinical state of ALS patients, type of ALS onset, or the duration of the disease. Decreased levels of CSF cGMP observed in this study may suggest the role of cGMP in neurodegeneration in ALS. The CSF cGMP cannot be a marker of the disease activity.     

The physician-patient relationship in amyotrophic lateral sclerosis

The principal models of the physician-patient relationship are analysed in terms of their historical development. An outline is given of the clinical, psychological and ethical particularities of the approach to patients with amyotrophic lateral sclerosis. The peculiarities of this disease are so exclusive that they do not resemble other progressive diseases with a negative prognosis, and therefore require an equally exclusive approach to the physician-patient relationship. This approach should not only be informative, scientific and interpretative-deliberative, but most simultaneously be founded on a solid therapeutic alliance aimed at seeking the best interests of the patients while respecting their autonomy as well as their “good” (not only in the sense of physical well-being, but also in terms of respect for their personal values). This is the only way to confront the conflicts that inevitably arise (especially in advanced stages of the disease) without the risks associated with a desire to escape or to adopt extreme solutions (such as euthanasia and therapeutic insistence) and without the risk of burn-out. Received: 5 May 2000 / Accepted in revised form: 6 December 2000     

Glutamate levels in cerebrospinal fluid in amyotrophic lateral sclerosis: a reappraisal using a new HPLC method with coulometric detection in a large cohort of patients.

Glutamate is involved in the degeneration of motor neurons in amyotrophic lateral sclerosis (ALS). However, the aetiology of ALS appears heterogeneous, leading to the possibility that patient subgroups with different pathophysiology may exist. The concentration of glutamate in cerebrospinal fluid (CSF) is measured using a new HPLC method with coulometric detection in a large cohort of ALS patients and controls: 377 ALS patients, 88 neurological patients and 18 normal controls. In ALS patients, and only in these subjects, the existence of two groups was observed, one with normal glutamate concentrations and one (40.8% of ALS patients) with high glutamate concentrations. High glutamate concentrations were correlated with a spinal onset of the disease, more impaired limb function and a higher rate of muscle deterioration. These results suggest that elevations of CSF glutamate concentrations could reflect the intensity of cell insult in the spinal cord. It remains to be determined if the group of patients with high CSF glutamate concentrations represents a specific subgroup of patients in terms of mechanism of disease, or only in terms of the spatial extent of motor neuron insult.     

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.     

Cerebrospinal fluid tau protein is not a biological marker in amyotrophic lateral sclerosis

Background:  Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder leading to progressive motor neuron cell death. Etiopathogenesis is still imperfectly known and much effort have been undertaken to find a biological marker that could help in the early diagnosis and in the monitoring of disease progression. Cerebrospinal fluid (CSF) concentrations of tau , an axonal microtubule-associated protein, have been measured in ALS with levels found increased in some studies and unchanged in others.
Methods:  Total CSF tau level was assayed in a population of ALS patients ( n =  57) and controls ( n =  110) using a specific ELISA method.
Results:  No significant differences in the median CSF tau levels between ALS cases and controls were found [ALS: 126 pg/ml (78–222); controls: 112 pg/ml (71–188), P =  ns]. In the ALS group, the bulbar-onset patients showed increased CSF tau levels as compared with the spinal-onset cases. These differences might be related to the higher age of the bulbar-onset patients. Further, no correlations were found between CSF tau concentrations and the rate of progression of the disease.
Conclusions:  These results do not support the hypothesis that total CSF tau protein is a reliable biological marker for ALS.     

Biochemical findings in multiple sclerosis

Summary Despite enormous efforts to find a specific laboratory test for multiple sclerosis, agar gel electrophoresis of the CSF proteins has remained the next to the best one. This study presents evidence that thin layer polyacrylamide isoelectric focusing of the CSF gamma globulins is by far superior for this purpose. In effect, about 91% of the 262 multiple sclerosis patients studied had oligoclonal fractions present in the very alkaline region of the pH gradient. Of the same group of patients, only 65% did show pathological results when studied by agar gel electrophoresis. Of the 272 CSF samples from patients suffering from other neurological diseases, only about 7% showed the presence of oligoclonal bands in the same region of the pH gradient, when submitted to isoelectric focusing.

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Zusammenfassung Unter den verschiedenen Laboratoriumstesten für multiple Sklerose war bisher die Agargel-Elektrophorese der Liquoreiweißfraktionen der Test mit dem höchsten diagnostischen Wert. In der vorliegenden Studie wird nachgewiesen, daß die isoelektrische Fokalisation der Gammaglobuline mittels Dünnschichtpolyacrylamidgel bei Multipler Sklerose weitaus wertvollere Ergebnisse zeitigt. Es ließen sich bei 91% von 262 Multiple-Sklerose-Patienten im alkalischen Bereich des pH-Gradienten oligoklonale Fraktionen im Liquor nachweisen. Bei der gleichen Patienten-Gruppe zeigten bei der Agargel-Elektrophorese nur 65% pathologische Ergebnisse. Unter 272 Liquorproben von Patienten, die an anderen neurologischen Erkrankungen litten, war nur bei 7% das Vorhandensein von oligoklonalen Zonen in der gleichen Region der pH-Gradienten bei isoelektrischem Fokusieren nachweisbar.

     

Creatine kinase activity in amyotrophic lateral sclerosis patients

Abstract. The aim of this study was to investigate creatine kinase (CK) in the serum of amyotrophic lateral sclerosis (ALS) patients. Previous investigations have shown an increased CK activity in ALS patients and this has been suggested to be an indicator of patients survival. The study was conducted at the Department of Neurology, University School of Medicine in Lublin. Thirty ALS patients took part in the study. The average duration of the disease was 17 months. Serum CK levels were measured by the enzymatic method with N-acethylcysteine. CK was elevated in 43.3% of the ALS patients. There were no significant differences in the serum CK level between the groups of the ALS patients depending on age, sex, duration of the disease, or clinical condition of patients. The CK level was significantly higher in the serum of the patients with a limb onset than in patients with a bulbar onset of ALS. Our study confirmed the increase in the serum CK activity in ALS patients. CK activity depends on a limb onset or a bulbar onset of ALS, but not on the duration of the disease and the severity of the clinical condition.     

Novel drug development for amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis (ALS) has become an increasingly attractive area for the pharmaceutical industry, the most experimentally tractable of the neurodegenerative diseases. Mechanisms underlying cell death in ALS are likely to be important in more common but more complex disorders. Riluzole, the only drug launched for treatment ALS is currently undergoing industrial trials for Alzheimer’s, Parkinson’s, Huntington disease, stroke and head injury. Other compounds in Phase III testing for ALS (mecamserin, xaliproden, gabapentin) are also in trials for other neurodegenerative disorders. Mechanisms of action of these advanced compounds are limited to glutamate antagonism, direct or indirect growth factor activity, as well as GABA agonism and interaction with calcium channels. A broader range of mechanisms is represented by compounds in Phase I trials: glutamate antagonism (dextramethorphan/p450 inhibitor; talampanel), growth factors (leukemia inhibiting factor; IL-1 receptor; encapsulated cells secreting CNTF) and antioxidants (TR500, a glutathione-repleting agent; recombinant superoxide dismutase; procysteine.) An even broader range of mechanisms is being explored in preclinical discovery programs. Recognition of the difficulties associated with delivery of protein therapeutics to the CNS has led to development of small molecules interacting either with neurotrophin receptors or with downstream intracellular signalling pathways. Other novel drug targets include caspaces, protein kinases and other molecules influencing apoptosis. High-throughput screens of large libraries of small molecules yield lead compounds that are subsequently optimized by chemists, screened for toxicity, and validated before a candidate is selected for clinical trials. The net is cast wide in early discovery efforts, only about 1% of which result in useful drugs at the end of a decade-long process. Successful discovery and development of novel drugs will increasingly depend on collaborative efforts between the academy and industry.