C-reactive protein in serum and cerebrospinal fluid in various neurological disorders: Apparent local consumption during bacterial meningitis

The level of C-reactive protein (CRP) was determined in the cerebrospinal fluid (CSF) by particle counting immunoassay. In non-neurological patients (N = 24), CRP was detectable only in 10 samples at concentrations ranging from 1.5 to 37 μg/l. The multiple sclerosis group did not differ from the controls. The highest CRP levels were found in viral and bacterial, including tuberculous, infections of the nervous system, with overlapping results for the various types of infections. However, in serum, the levels of CRP were much higher in pyogenic than in viral meningitis. We compared the CSF CRP/serum CRP ratio to the same ratio for albumin and found a significant correlation between the two ratios in viral, but not in bacterial, infections. These results suggest a local consumption of CRP during bacterial meningitis.     

Differentiation of the humoral immune response in inflammatory diseases of the central nervous system

Summary The three main immunoglobulin classes obey the basic principles of passive protein transfer at the blood-CSF barrier and the serum-derived portions could therefore be quantified with the help of the permeability marker albumin. The Ig fractions secreted into the CSF by sessile plasma cell clones have been determined in various inflammatory diseases of the central nervous system. The humoral immune response in multiple sclerosis and chronic encephalitis of unknown cause was dominated by IgG antibodies. In most other inflammatory diseases IgA and IgM were concomitantly synthesized, e.g. in neurosyphilis and meningoencephalitis caused by viruses of the herpes group. In tick-borne meningopolyneuritis Bannwarth, only IgM and in bacterial meningitis only IgA may be produced locally. The detection of a secretory immunoglobulin fraction in the CSF may be the sole laboratory parameter in chronic inflammatory processes of the nervous system.

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Zusammenfassung Die drei Immunglobulin-Klassen G, A und M befolgen die Gesetze des passiven Proteintransfers an der Blut-Liquor-Schranke. Deshalb können die aus dem Serum stammenden Anteile mit Hilfe des Permeabilitäts-markers Albumin quantifiziert werden. Die IgG-Fraktionen, die durch gewebsständige Plasmazell-Klone bei verschiedenen entzündlichen Erkrankungen des Zentralnervensystems in den Liquor abgegeben werden, konnten quantitativ bestimmt werden. Bei der multiplen Sklerose und der chronischen Encephalitis ungeklärter Ursache wird überwiegend IgG synthetisiert. Bei den meisten anderen entzündlichen Erkrankungen wird zur gleichen Zeit auch IgA und IgM gebildet, etwa bei der Neurolues und der Herpes-Encephalitis. Bei der durch Zecken übertragenen Meningo-Polyneuritis Bannwarth wird gelegentlich nur IgM und bei der bakteriellen Meningitis nur IgA synthetisiert. Eine lokal gebildete Immunglobulin-Fraktion kann der einzige klinisch-chemische Parameter für das Vorliegen einer chronisch-entzündlichen Erkrankung des Nervensystems sein.

     

The polymerase chain reaction and its applications in neuropathology

The polymerase chain reaction (PCR) provides a means of generating large numbers of copies of selected segments of DNA. Once amplified, the DNA can be characterized by determination of its size and sequence. For many applications, routinely-processed biopsy and autopsy material is an adequate substrate for the reaction. PCR can be used to amplify sequences of DNA that are uniquely characteristic of particular micro-organisms, allowing their rapid detection in samples of tissue or cerebrospinal fluid. The technique allows clones of cells with gene rearrangements or translocations to be detected with great sensitivity and is proving a useful way to monitor the effects of tumour therapy, particularly in patients with lymphomas and leukaemias. Further applications include the identification of gene deletions and mutations, and the assessment of cell lineage by amplification and analysis of highly polymorphic gene loci (DNA fingerprinting). Because the degree of amplification resulting from PCR is so great, even a single molecule of contaminating DNa may be detected and its significance misinterpreted. Great care, therefore, should be taken to prevent contamination of samples, particularly by the products of previous reactions, and every series of reactions should include appropriate controls.     

脑脊液细胞学在中枢神经系统白血病诊治中的应用

目的探讨脑脊液细胞学在中枢神经系统白血病(CNSL)的诊断、病情进展及指导治疗中的作用。方法采用粟氏FMU-5型细胞玻片离心沉淀仪收集脑脊液细胞,迈-格-姬(MGG)染色,显微镜下进行细胞分类,计数白血病细胞所占的百分比。分析2000～2008年我院住院的58例CNSL患者的脑脊液细胞学检查结果。结果 58例患者包括19例儿童和39例成人病例。其脑脊液细胞学均发现白血病细胞,占脑脊液细胞的5～96%。其中5例首次检测诊断为CNSL后失访,其余病例均于治疗过程中重复进行脑脊液细胞学检查。白血病细胞所占比例多数呈不同程度的降低,直至完全消失;11例比值出现反复,最终逐渐降低;8例未见明显降低。结论脑脊液细胞学检查对CNSL的诊断、治疗及预后评估具有重要的临床意义。     

Cerebrospinal fluid lavage in the treatment of inadvertent intrathecal vincristine injection

Vincristine, a widely used antineoplastic agent, is extremely toxic to the central nervous system. If given intrathecally, it produces a rapidly ascending, usually fatal, neuromyeloencephalopathy. We report a case of this complication in a 7-year-old girl with acute lymphoblastic leukemia who was receiving maintenance chemotherapy. During one treatment 0.5 mg of vincristine was erroneously injected into the lumbar subarachnoid space. Cerebrospinal fluid lavage was established within 2 h and continued for 24 h. After 7 days she developed a progressive sensorimotor paraplegia, which eventually stabilized as a paraparesis. Neurophysiological studies were consistent with an axonal type sensorimotor neuropathy. Magnetic resonance imaging of the spine was normal. Vincristine binds to cells, blocking mitosis, thus causing cell death. The associated central nervous system lesions are those of an ascending chemical leptomeningitis and ventriculitis. Cerebrospinal fluid lavage dilutes and removes the drug, thus limiting neural damage. At present this is the only treatment for intrathecal vincristine injection, and its early use in such an event is considered mandatory. Received: 10 June 1998     

Polyclonal and oligoclonal IgA synthesis in the cerebrospinal fluid of neurological patients: An immunoaffinity-mediated capillary blot study

An intrathecal synthesis of IgA has been reported in various neurological disorders. However, the frequency of its occurence and the electrophoretic characteristics of the locally produced IgA remained a matter of controversy. We developed a sensitive immunoaffinity-mediated capillary blot technique for the detection of polyclonal and oligoclonal IgA in the CSF of 115 patients with various neurological disorders. Paired CSF and serum samples containing 50 ng IgA after appropriate dilutions were submitted to isoelectric focusing in agarose gels; IgA was then blotted onto a polyvinylidene difluoride sheet coated by an anti-IgA antiserum or by infectious antigens. The immunoblots were revealed by an alkaline phosphatase-conjugated anti-IgA antiserum. Only five samples displayed CSF-restricted oligoclonal IgA bands, including two out of 33 from MS patients. In herpetic encephalitis (n = 5) and varicella-zoster meningitis (n = 2), a strong intrathecal production of virus-specific IgA antibodies was detectable. In such cases, faint oligoclonal IgA antibodies were superimposed on a polyclonal background. A weak local production of anti-Borreliaburgdorferi IgA antibodies was present in two out of four cases of neuroborreliosis.     

Monitoring and tentative diagnosis of herpetic encephalitis by protein analysis of cerebrospinal fluid. Particular relevance of the assays of ferritin and S-100

The cerebrospinal fluid (CSF) of 9 patients with herpetic encephalitis was analyzed by particle counting immunoassay of ferritin, S-100, immunoglobulins, anti-herpes antibodies and immune complexes and by electrophoresis for the detection of oligoclonal bands. The main conclusions are: first, the simultaneous increase of both ferritin and S-100 in the presence of symptoms of encephalitis suggests strongly the infection is herpetic; second, high and increasing levels of S-100, probably related to the extent of the necrotic process, indicate a poor prognosis. In addition, 8-14 days after onset, locally produced anti-herpes antibodies were detectable, the IgG index increased and oligoclonal bands became visible.     

Myoglobin as a polymerase chain reaction (PCR) inhibitor: A limitation for PCR from skeletal muscle tissue avoided by the use of thermus thermophilus polymerase

Skeletal muscle tissue contains polymerase chain reaction (PCR) inhibitors that are coextracted by conventional nucleic acid extraction procedures. Myoglobin, a heme-containing molecule, was shown to act as a potent Thermus aquaticus DNA polymerase inhibitor and is likely to be involved in muscle tissue-associated PCR inhibition. The use of Thermus thermophilus DNA polymerase avoids muscle tissue-associated PCR inhibition, and should be used in case of small amounts or instability of the targeted nucleic acid. © 1998 John Wiley & Sons, Inc. Muscle Nerve 21:1064–1067, 1998.     

笑气滥用导致神经功能缺损的临床特点

目的 探讨笑气滥用导致神经功能缺损的临床特点.方法 回顾性分析2例笑气滥用导致神经功能缺损患者的临床资料,并对相关文献资料进行复习.结果 1例以模拟格林-巴利综合征的症状发病,表现为四肢对称性的麻木无力,脊椎横断面T2WI显示颈髓后索倒"V"形高信号,EMG呈多发性周围神经脱髓鞘和轴索损伤.另1例以模拟急性脊髓炎症状起...     

Adenine nucleotide metabolism and cell fate after oxidant exposure of rat cortical neurons: effects of inhibition of poly(ADP-ribose) polymerase

We exposed cultured neurons prelabeled with 14C-adenine to H2O2 with or without the poly(ADP-ribose) polymerase (PARP) inhibitor 3,4-Dihydro-5-[4-(1-piperidinyl)butoxy]-1(2H)-isoquinolinone (DPQ) to quantify its effects on acute ATP depletion, later ATP synthesis, cellular and nuclear morphology, extent of DNA fragmentation, and PARP cleavage. According to the extent of the acute ATP depletion, the exposures were classified as 'mild' (50 microM H2O2), 'moderate' (100-250 microM H2O2), or 'severe' (500 microM-1 mM H2O2) insults. Mild exposure had no significant effects on the parameters studied. In the 'moderately' exposed neurons, ATP depletion to 59+/-6% of control was associated with a decrease in the cell counts, apoptotic morphology, and cleavage of PARP. In this group, DPQ prevented the acute ATP (to 95+/-15% of control), preserved cell morphology, and improved cell survival. In the 'severe' group, ATP depletion to 18+/-4% was associated with necrosis and intact PARP. DPQ elevated ATP levels (to 44+/-12% of control) and post-insult ATP synthesis, improved cell counts, and altered cell morphology towards apoptosis rather than necrosis. Post-insult application of DPQ was less effective. Our results show that the extent of oxidant-induced ATP depletion and cell fate can be modified by PARP inhibition, to some extent also after the insult.     

Beneficial effects of PJ34 and INO-1001, two novel water-soluble poly(ADP-ribose) polymerase inhibitors, on the consequences of traumatic brain injury in rat

Traumatic brain injury produces peroxynitrite, a powerful oxidant which triggers DNA strand breaks, leading to the activation of poly(ADP-ribose)polymerase-1 (PARP-1). We previously demonstrated that 3-aminobenzamide, a PARP inhibitor, is neuroprotective in a model of traumatic brain injury induced by fluid percussion in rat, suggesting that PARP-1 could be a therapeutic target. In order to confirm this hypothesis, we investigated the effects of PJ34 and INO-1001, two PARP inhibitors from structural classes other than benzamide, on the post-traumatic consequences. Pre- and post-treatments with PJ34 (30 mg/kg/day) and INO-1001 (10 mg/kg/day) decrease the neurological deficit at 3 days post-injury and this deficit is still reduced at 7 days. These neurological recovery-promoting effects are associated with the inhibition of PARP-1 activation caused by trauma, as demonstrated by abolishment of immunostaining of poly(ADP-ribose). Thus, the present work strengthens strongly the concept that PARP-1 inhibition may be a suitable approach for the treatment of brain trauma.     

Immunoglobulin M specific for measles in serum and cerebrospinal fluid of patients with multiple sclerosis and other neurological diseases

Summary Serum and cerebrospinal fluid of patients with multiple sclerosis, subacute sclerosing panencephalitis and other neurological diseases have been tested by the indirect fluorescent antibody method for immunoglobulin M specific for measles. Only sera of three patients were positive. This feature is of little statistical importance. Nevertheless the authors emphasize the role of a possible viral infection in the pathogenesis of multiple sclerosis.Presented to the 12th Meeting of the Section of Neuropathology of the Italian Society of Neurology, Massa Marittima, June 5–6, 1976     

磁共振成像及脑脊液细胞学对病毒性脑(膜)炎的诊断性研究

目的探讨MRI和脑脊液细胞学检测阳性率、异常程度和发病时间对病毒性脑(膜)炎诊断和制定治疗方案的指导价值。方法回顾189例病毒性脑(膜)炎患者的头部MRI及脑脊液检查资料,分析MRI正常或异常患者在疾病早期的脑脊液细胞学改变。结果 189例患者中96例(50.79%)呈现MRI异常影像、129例(68.25%)脑脊液细胞学检测异常。MRI异常患者中脑脊液细胞学检测异常率为72.92%(70/96),与MRI正常患者(63.44%,59/93)比较差异具有统计学意义(P=0.000)。结论病毒性脑(膜)炎患者在疾病早期MRI改变晚于脑脊液细胞学改变,但二者对提示诊断和制定治疗方案均具有重要临床意义。     

两步—多重PCR诊断134例假肥大型进行性肌营养不良症基因缺失

研究临床检测假肥大型进行性肌营养不良症（ＤＭＤ／ＢＭＤ）基因缺失的有效手段。方法运用两步－多重聚合酶链反应（ＰＣＲ）技术诊断ＤＭＤ／ＢＭＤ基因缺失。结果１３４例病人中，检测阳性率为４９．３％（６６／１３４）。结论多重ＰＣＲ诊断ＤＭＤ／ＢＭＤ基因缺失敏感、快速、准确，可在临床推广应用。     

Multivariate analysis of the serum-cerebrospinal fluid-protein-relation for the diagnosis of neurological diseases of the central nervous systems

ABSTRACT In a population comprising 197 patients, serum and CSF proteins were assayed using the radial immunodiffusion technique devised by Mancini. Multiple discriminants analysis was applied to investigate whether the measured CSF/serum protein relations and their ratios could be regarded as an indicator of specific neurological diseases. One significant finding was that the slope angle α of the regression line between the serum/CSF relation and molecular weight may represent an important indicative parameter. A small angle is suggestive of enhanced permeability of the BBB, a large angle of a correspondingly lowered permeability. Further, the analyses demonstrated that the combined use of several predictors can markedly improve differential diagnosis. The study also demonstrates the potential of a statistical analytic technique that is still rarely applied in medicine.     

Alteration of BAFF and APRIL in the cerebrospinal fluid based on the therapeutic response in primary central nervous system B-cell lymphoma

We evaluated the cerebrospinal fluid (CSF) levels of the B-cell activating factor of the tumor necrosis factor family (BAFF) and A proliferation-inducing ligand (APRIL) in two cases of primary central nervous system B-cell lymphoma (PCNSBL) before and after treatment. One patient achieved clinical remission, and demonstrated decrease in the CSF levels of both BAFF and APRIL after treatment. Meanwhile, the other patient with insufficient therapeutic response showed increase in the BAFF levels despite decrease in APRIL levels. This report suggests that the combination of BAFF and APRIL levels could be useful in estimating the therapeutic efficacy in treating PCNSBL as reliable CSF markers.     

Saturable uptake of [I] -triiodothyronine at the basolateral (blood) and apical (cerebrospinal fluid) sides of the isolated perfused sheep choroid plexus

The uptake of ¹²⁵I-labelled -triiodothyronine (T₃) was measured on the blood side of the isolated perfused choroid plexus of the sheep using steady-state and single-circulation paired tracer techniques. The steady-stake uptake of T₃ was 33.5% (perfussion fluid protein content was 0.05 g·dl⁻¹) which could be reduced to 9.4% in the presence of 500 μM unlabelled T₃ showing partial saturation. The CSF to blood steady-state [¹²⁵I]T₃ measurements gave plasma/CSF ratio, R%, of 24.6 ± 4.8% which was reduced to 9.8 ± 2.1% in the presence of 500 μM unlabelled T₃ in the mock CSF. The transport of T₃ across the blood face of the choroid plexus and the CSF to blood transport, failed to show sodium dependence. Using the single circulation paired tracer technique, the initial uptake in less than 60 s, U_max of [¹²⁵I]T₃ was 50.4 ± 3.9% relative to the extracellular marker [³H] -mannitol. However, when 250 μM unlabelled T₃ was present, U_max was reduced by 66%, although further significant inhibition at higher concentrations was not observed. Uptake of T₃ at the blood side of the choroid plexus was partially saturated in the presence of unlabelled reverse T₃ and DT₃, suggesting little uptake stereospecificity. Unlabelled thyroxine (T₄) and the amino acid analogues cycloleucine (aminocyclopentane-1-carboxylic acid) and BCH (β-2-aminobicyclo-[2,2,1]-heptane-2-carboxylic acid) each reduced [¹²⁵I]T₃ uptake significantly, but not to the same degree as T₃ stereoisomers. The neutral amino acids alanine and phenylalanine, had no effect on uptake. The [¹²⁵I]T₃ unidirectional flux was calculated from U_max vlues and the resultant kinetic curve could be resolved into two components; a non-saturable process with a slope of 1.2 ml·min⁻¹·g⁻¹, and a saturable process with K_m = 66 ± 22 μM and V_max = 0.44 ± 0.11 μmol·min⁻¹·g⁻¹. These data suggest that [¹²⁵I]T₃ uptake at the blood face of the choroid plexus, is mediated by both saturable and non-saturable uptake processes, which lack stereospecificity, sodium dependence, and exhibit cross competition both with T₄ and the large neutral amino acid analogues, cycloleucine and BCH. Transport from CSF was also partially saturable, did not exhibit sodium dependence.     

Elevation of Gas6 protein concentration in cerebrospinal fluid of patients with chronic inflammatory demyelinating polyneuropathy (CIDP)

INTRODUCTION: Gas6 enhances survival of Schwann cells and neurons in vitro and participates in autoimmunity in animal models. Since its concentration in human cerebrospinal fluid (CSF) is unknown, we measured it in samples from patients with non-inflammatory/non-autoimmune neurological diseases (NINAD) and autoimmune polyneuropathies. MATERIALS AND METHODS: Samples collected after informed consent during diagnostic lumbar puncture in the period 1999-2006 were stored at -30 degrees C. We considered subjects with NINAD (stroke, ALS, headache, psychiatric conditions simulating neurological diseases, otologic dizziness) or with Guillain-Barré syndrome (GBS) or CIDP. CSF and plasma total protein and age were obtained from clinical records. Gas6 was measured with an ELISA developed and validated in our laboratory (inter-, intra-assay CVs <10%, recovery 96%). Variance, Tukey's post-hoc test, regression were calculated with a statistical software (Statsoft). RESULTS: Mean Gas6 concentration in patients with NINAD was 6.5+/-2.4 ng/ml, 7.2+/-2.6 ng/ml in GBS and significantly higher (11.5+/-1.7 ng/ml) in CIDP than in the other conditions (post-hoc, p<0.005). It was not related to age, CSF total proteins or to CSF/plasma ratio of total proteins (regression, p>0.1). CONCLUSIONS: Gas6 is detectable in CSF and may be involved in chronic autoimmune demyelination or myelin repair.