Serum procalcitonin and cerebrospinal fluid cytokines level in children with meningitis

AIMS: To determine the level of serum procalcitonin and cerebrospinal fluid cytokines in children with bacterial or viral meningitis and to document the use of these parameters in differential diagnosis. RESULTS: Before the start of antibiotic treatment, serum procalcitonin and tumor necrosis factor alpha levels were found to be higher in acute bacterial meningitis compared with viral meningitis and with the control group. Similarly, cerebrospinal fluid interleukin-6 levels were found to be significantly higher in children with acute bacterial meningitis compared with viral meningitis. However, no significant difference was determined between groups in respect to the cerebrospinal fluid interleukin-8 level. CONCLUSION: Serum procalcitonin and cerebrospinal fluid tumor necrosis factor alpha levels can be used in the early diagnosis of bacterial meningitis. Similarly, they may be useful adjuncts in differential diagnosis of bacterial and viral meningitis.     

醒脑静注射液联合血府逐瘀汤对SAH大鼠CRP、TNF-alpha的影响

目的：探讨醒脑静注射液联合血府逐瘀汤对蛛网膜下腔出血（subarachnoidhemorrhage,SAH）大鼠C反应蛋白(C-reactionprotein, CRP) 、肿瘤坏死因子-a（Tumor necrosis factor-a,TNF-a）的影响。方法：50 只大鼠随机分为5 组：正常组,模型组,醒脑静组 （0.3 mL/kg）,血府逐瘀汤组（0.3 mL/kg）,联合用药组（醒脑静联合血府逐瘀汤）。除正常组外,其余4组采用" 二次枕大池注血法" 建立大鼠SAH 模型。从造模成功后的第1 d 开始给药,连续给药7 d。正常对照组及模型组给予等量生理盐水。分别在术前、术后 1 d、3 d、7 d、14 d取血浆及脑脊液,按ELISA 试剂盒说明书分别测定各组血清及脑脊液中肿瘤坏死因子-alpha（TNF-alpha）、C- 反应蛋白 （CRP）的含量。结果：（1）术后随着时间推移,SAH大鼠血清及脑脊液中TNF-alpha及CRP 含量逐渐上升,在第7 d 达到高峰。（2）术 后第1 d,与正常组比较,模型组中血清及脑脊液中TNF-琢、CRP含量均显著上升,差异显著（P<0.01）;与模型组比较,醒脑静组及 血府逐瘀汤组中血清及脑脊液中TNF-alpha、CRP 含量无差异（P>0.05）,而联合用药组中血清及脑脊液中TNF-琢含量下降,差异具有 显著意义（P<0.05）。（3）术后第3 d、7 d、14 d,与正常组比较,模型组中血清及脑脊液中TNF-琢、CRP 含量均显著上升,差异显著 （P<0.01）;与模型组比较,醒脑静组、血府逐瘀汤组及联合用药组中血清及脑脊液中TNF-琢、CRP 含量下降,差异均显著（P<0.0 1）,与醒脑静组或血府逐瘀汤组比较,联合用药组中血清及脑脊液中TNF-alpha、CRP含量下降,差异均显著（P<0.05）。结论：醒脑静 联合血府逐瘀汤能有效缓解SAH大鼠CVS,与降低SAH大鼠血清及脑脊中TNF-alpha、CRP 水平有关。     

结核性脑膜炎与化脓性脑膜炎脑脊液与血浆生化指标比值的比较研究

目的:比较结核性脑膜炎与化脓性脑膜炎脑脊液与血浆生化指标比值。方法:选择2010年2月~2014年12月我院结核性脑膜炎患者82例,化脓性脑膜炎98例,检测脑脊液与血浆中的蛋白、糖及氯化物含量,并计算比值。结果:两组患者脑脊液蛋白、糖含量的差异无统计学意义(P0.05),化脓性脑膜炎组的氯化物含量高于结核性脑膜炎组(P0.05);两组血浆糖含量的差异无统计学意义(P0.05),化脓性脑膜炎组蛋白和氯化物含量明显高于结核性脑膜炎组(P0.05);化脓性脑膜炎组蛋白比值低于结核性脑膜炎组,氯化物比值则高于化脓性脑膜炎组,差异均有统计学意义(P0.05);两组间糖比值比较,差异无统计学意义(P0.05)。结论:脑脊液与血浆生化指标比值对鉴别诊断结核性脑膜炎与化脓性脑膜炎有重要意义。     

中枢神经系统感染患儿血清和脑脊液CRP、PCT、TNF-α及MMP-9水平及其临床意义

目的:探讨中枢神经系统感染患儿血清和脑脊液C反应蛋白(CRP)、降钙素原(PCT)、肿瘤坏死因子-α(TNF-α)及基质金属蛋白酶-9(MMP-9)水平及其临床意义。方法:选择2017年1月~2018年6月期间南京市第二医院收治的中枢神经系统感染患儿93例作为研究对象,其中化脓性脑膜炎62例记为化脓性脑膜炎组,病毒性脑炎31例记为病毒性脑炎组,另选取同期于我院治疗的非中枢神经系统感染患儿40例作为对照组,比较各组血清、脑脊液CRP、PCT、TNF-α、MMP-9水平及阳性率,并计算血清和脑脊液CRP、PCT、TNF-α、MMP-9诊断中枢神经系统感染的灵敏度、特异度及准确度。结果:化脓性脑膜炎组患儿血清、脑脊液CRP、PCT、TNF-α及MMP-9水平及阳性率高于病毒性脑炎组和对照组,病毒性脑炎组患儿血清、脑脊液CRP、TNF-α及MMP-9水平及阳性率高于对照组(P<0.05),病毒性脑炎组与对照组血清、脑脊液PCT水平及阳性率比较无统计学差异(P>0.05)。血清或脑脊液CRP+PCT+TNF-α+MMP-9联合检验对中枢神经系统感染具有一定的诊断价值。结论:中枢神经系统感染患儿血清、脑脊液CRP、TNF-α、PCT及MMP-9水平明显升高,其中化脓性脑膜炎患儿血清、脑脊液PCT水平高于病毒性脑炎患儿,血清或脑脊液CRP、PCT、TNF-α及MMP-9联合检验对儿童中枢神经系统感染的鉴别诊断具有较高的价值。     

Gonadotropin and prolactin secretion following intraventricular administration of morphine in gilts

The effects of central nervous system administration of morphine on secretion of luteinizing hormone (LH), follicle-stimulating hormone, and prolactin were investigated in ovariectomized gilts stereotaxically implanted with lateral ventricular cannulas. In Experiment 1, mean serum LH and follicle-stimulating hormone concentrations and serum LH pulse frequency were unaffected by artificial cerebrospinal fluid administration (P greater than 0.1), but decreased (P less than 0.01) in 8 of 11 gilts when 500 micrograms of morphine were given 3 hr later. Serum LH pulse amplitude was unaffected (P greater than 0.1) by cerebrospinal fluid or morphine injection. In Experiment 2, luteinizing hormone concentrations decreased (P less than 0.0001) and prolactin concentrations increased (P less than 0.0001), but follicle-stimulating hormone concentrations did not change (P greater than 0.1) after 500 micrograms of morphine. Gonadotropin responses to 10 micrograms of gonadotropin-releasing hormone, given 2 hr after intraventricular injection, were similar (P greater than 0.1) for morphine- and cerebrospinal fluid-treated gilts. These results indicate that morphine inhibits LH secretion at the level of the central nervous system, and are consistent with the concept that endogenous opioid peptides participate in the regulation of gonadotropin and prolactin release in pigs.     

The effect of ACTH4-9 analog (Org2766) on some cerebrospinal fluid parameters in patients with Alzheimer's disease

In a double-blind, placebo-controlled study, the central nervous system effects of an ACTH4-9 analog, Org2766 (40 mg/day), in Alzheimer's disease were assessed by measuring cerebrospinal fluid parameters during 6 months' treatment. Somatostatin-like immunoreactivity and cholinesterase activity, which are known to be reduced in cerebrospinal fluid of Alzheimer patients compared with controls, did not change during treatment. As a marker of noradrenergic and dopaminergic systems, we measured dopamine-beta-hydroxylase and homovanillic acid, but both levels were static. These results suggest that Org2766 did not interact with the transmitter systems, which are thought to be disturbed in Alzheimer's disease.     

Turbidimetric determination of lysozyme with Micrococcus lysodeikticus cells: Reexamination of reaction conditions

Factors affecting the activity of human lysozyme (EC 3.2.1.17) toward cell suspensions of Micrococcus lysodeikticus were reexamined. Effects of substrate concentration, pH, and ionic strength and matrix effects of protein were assessed with special emphasis on the interdependence of various parameters. On the basis of these evaluations, an optimized kinetic turbidimetric method for lysozyme assay was set up. The method was applied for automation with a System Olli 3000 analyzer. The new automated lysozyme assay proved good for routine clinical use in regard to analysis speed, sensitivity, linearity, and reproducibility. Reference values for serum, urinary, and cerebrospinal fluid lysozyme were assessed with the automated method.     

脑脊液乳酸和血清降钙素原及C反应蛋白 对小儿细菌性脑膜炎的诊断价值

目的探讨脑脊液乳酸、血清降钙素原及C反应蛋白对小儿细菌性脑膜炎的诊断价值。方法选取我院2016年4月至2017年6月收治的50例细菌性脑膜炎患儿以及50例病毒性脑膜炎患儿进行作为研究对象,比较2类患儿脑脊液乳酸(LA)、血清降钙素原(PCT)及C反应蛋白(CRP)的水平,并分析其诊断价值。结果细菌性脑膜炎组患儿脑脊液LA、血清PCT及CRP水平显著高于病毒性脑膜炎患儿(均P0.05)。血清PCT诊断的灵敏度和特异度最高(96.4%、90.9%,P0.05)。3项指标联合检测的灵敏度(100.0%)和特异度(95.5%)明显高于任一单项指标(均P0.05)。经过Pearson相关性分析,脑脊液LA、血清PCT及CRP与小儿细菌性脑膜炎均呈显著正相关关系(均P0.05)。结论脑脊液乳酸、血清PCT及CRP对小儿细菌性脑膜炎的诊断和治疗效果监测有重要应用价值。     

The effects of serum osmolarity on cerebrospinal fluid volume flow

The effects of changes in serum osmolarity on cerebrospinal fluid (CSF) formation were studied in cats. CSF production rates were measured by ventriculocisternal perfusion both before and after intravenous infusion of glucose solutions.Infusion of glucose, hyperosmolar with respect to serum, increased serum osmolarity and caused a decrease in CSF formation rate; conversely, infusion of hypoosmolar solutions lowered serum osmolarity and increased CSF formation. CSF production and serum osmolarity were found to be linearly related. A 1% serum osmolarity change resulted in a 6.7% change in CSF formation. CSF formation increased by 130% with a serum osmolarity of 265 m0sm/1 and was undetectable with serum of 380 m0sm/1.     

Horses experimentally infected with Sarcocystis neurona develop altered immune responses in vitro

Equine protozoal myeloencephalitis (EPM) due to Sarcocystis neurona infection is 1 of the most common neurologic diseases in horses in the United States. The mechanisms by which most horses resist disease, as well as the possible mechanisms by which the immune system may be suppressed in horses that develop EPM, are not known. Therefore, the objectives of this study were to determine whether horses experimentally infected with S. neurona developed suppressed immune responses. Thirteen horses that were negative for S. neurona antibodies in serum and cerebrospinal fluid (CSF) were randomly assigned to control (n = 5) or infected (n = 8) treatment groups. Neurologic exams and cerebrospinal fluid analyses were performed prior to, and following, S. neurona infection. Prior to, and at multiple time points following infection, immune parameters were determined. All 8 S. neurona-infected horses developed clinical signs consistent with EPM, and had S. neurona antibodies in the serum and CSF. Both infected and control horses had increased percentages (P < 0.05) of B cells at 28 days postinfection. Infected horses had significantly decreased (P < 0.05) proliferation responses as measured by thymidine incorporation to nonspecific mitogens phorbol myristate acetate (PMA) and ionomycin (I) as soon as 2 days postinfection.     

Tropical spastic paraparesis/HTLV-I associated myelopathy

In 1985 we had the first indication that human T-cell lymphotropic virus (HTLV-I) was the possible etiological agent of a chronic myelopathy that seemed to be peculiar to the tropics and that is now known as endemic tropical spastic paraparesis (TSP). IgG antibodies to HTLV-I were found in serum and cerebrospinal fluid of patients from Jamaica, Colombia, Martinique, and shortly after in southern Japan, where the disease is called HTLV-I-associated myelopathy (HAM). The HTLV-I seropositivity was first determined by enzyme-linked immunoassay and confirmed by western immunoblot and in the cerebrospinal fluid specific IgG oligoclonal bands to HTLV-I were found in cerebrospinal fluid and not in serum. These laboratory findings indicated that HTLV-I could be neuropathogenic and for the first time a single etiological agent was identified in patients from different countries. Thus, in less than a decade a century of research and speculation was seemingly resolved when this disease, which was thought to occur only in blacks of poor socieconomic status in tropical countries, was shown to occur in all ethnic groups of varying socioeconomic status in temperate, subtropical, and tropical climates.     

CSF proteome: a protein repository for potential biomarker identification

Proteomic analysis is not limited to the analysis of serum or tissues. Synovial, peritoneal, pericardial and cerebrospinal fluid represent unique proteomes for disease diagnosis and prognosis. In particular, cerebrospinal fluid serves as a rich source of putative biomarkers that are not solely limited to neurologic disorders. Peptides, proteolytic fragments and antibodies are capable of crossing the blood-brain barrier, thus providing a repository of pathologic information. Proteomic technologies such as immunoblotting, isoelectric focusing, 2D gel electrophoresis and mass spectrometry have proven useful for deciphering this unique proteome. Cerebrospinal fluid proteins are generally less abundant than their corresponding serum counterparts, necessitating the development and use of sensitive analytical techniques. This review highlights some of the promising areas of cerebrospinal fluid proteomic research and their clinical applications.     

Invasive meningococcal disease and latex agglutination test — Is it still beneficial for diagnosis?

We showed current clinical usefulness of the latex agglutination (LA) test for confirmation of meningococcal etiology on 32 cerebrospinal fluid, 77 serum and 93 urine samples collected during the first week of hospitalization from 19 patients with laboratory confirmed invasive meningococcal disease. The positivity of the LA test in cerebrospinal fluid was 47%, in serum 42% and in urine 24%, while the PCR of cerebrospinal fluid and serum was positive in 95 and 47% cases, respectively. The latest positivity of the LA test was detected on day 2 in cerebrospinal fluid, on day 3 in serum and on day 4 in urine. In the group of patients who had received antibiotic therapy we found nonsignificant reduction of LA positivity and also statistically significant reduction of culture positivity in CSF (p = 0.04); the PCR positivity changed minimally. In blood samples, nonsignificant reduction of culture positivity and no difference in LA and PCR positivity was found. We did not find any statistically significant relationship between test results and clinical forms. The LA test can be therefore considered to be an auxiliary diagnostic method, rapid and easily practicable but less sensitive than PCR. It can be recommended especially for local laboratories where PCR is not available and the patient already received antibiotics before admission to the hospital.     

Profiles of Extracellular miRNA in Cerebrospinal Fluid and Serum from Patients with Alzheimer's and Parkinson's Diseases Correlate with Disease Status and Features of Pathology

The discovery and reliable detection of markers for neurodegenerative diseases have been complicated by the inaccessibility of the diseased tissue- such as the inability to biopsy or test tissue from the central nervous system directly. RNAs originating from hard to access tissues, such as neurons within the brain and spinal cord, have the potential to get to the periphery where they can be detected non-invasively. The formation and extracellular release of microvesicles and RNA binding proteins have been found to carry RNA from cells of the central nervous system to the periphery and protect the RNA from degradation. Extracellular miRNAs detectable in peripheral circulation can provide information about cellular changes associated with human health and disease. In order to associate miRNA signals present in cell-free peripheral biofluids with neurodegenerative disease status of patients with Alzheimer''s and Parkinson''s diseases, we assessed the miRNA content in cerebrospinal fluid and serum from postmortem subjects with full neuropathology evaluations. We profiled the miRNA content from 69 patients with Alzheimer''s disease, 67 with Parkinson''s disease and 78 neurologically normal controls using next generation small RNA sequencing (NGS). We report the average abundance of each detected miRNA in cerebrospinal fluid and in serum and describe 13 novel miRNAs that were identified. We correlated changes in miRNA expression with aspects of disease severity such as Braak stage, dementia status, plaque and tangle densities, and the presence and severity of Lewy body pathology. Many of the differentially expressed miRNAs detected in peripheral cell-free cerebrospinal fluid and serum were previously reported in the literature to be deregulated in brain tissue from patients with neurodegenerative disease. These data indicate that extracellular miRNAs detectable in the cerebrospinal fluid and serum are reflective of cell-based changes in pathology and can be used to assess disease progression and therapeutic efficacy.     

创伤后应激障碍大鼠海马与前额叶皮质中OREXIN受体的失调

目的:从食欲素(Orexin, ORX)系统挖掘创伤后应激障碍的神经生物学机制。方法:以单次延长刺激(single prolonged stimulation,SPS)法复制创伤后应激障碍(post traumatic stress disorder,PTSD)大鼠模型,以行为学结合血清中皮质酮和神经元特异性烯醇化酶进行模型评价,通过酶联免疫吸附试验(Elisa)分析大鼠血清与脑脊液中OREXIN水平,以实时荧光定量聚合酶链反应(RT-PCR)检测海马与前额叶皮质中的Orexin受体基因表达。结果:实验成功的复制了PTSD模型,与对照组相比,模型组大鼠血清中皮质酮和神经元特异性烯醇化酶的含量均极显著升高(P0.01),脑脊液中Orexin A、Orexin B的含量均显著升高(P0.01),海马和皮质中ORX1R与ORX2R均极显著下降(P0.01)。结论:PTSD大鼠的应激损伤与Orexin及其受体表达水平的变化密切相关。     

Assay of inorganic sulfate in biologic fluids by nonsuppressed (single-column) ion chromatography

An assay using nonsuppressed (single-column) anion chromatography was developed to determine the concentration of inorganic sulfate in biologic fluids. A conventional HPLC system with an anion-exchange column and conductimetric detector interfaced with an automatic injector and integrator was used. The mobile phase for the chromatography of urine and serum samples is 4 mM potassium hydrogen phthalate, pH 4.5, and potassium iodide is used as the internal standard. For cerebrospinal fluid samples, the mobile phase is modified by addition of 10% of a 4 mM phthalic acid solution. Results of the HPLC assay were found to correlate well (r = 0.991 and 0.999) with those of two commonly used spectrophotometric methods for urine and serum inorganic sulfate determinations. However, the concentrations determined by ion chromatography were 2.5 to 10% lower, possibly due to less assay interference by other substances following chromatographic separation of sulfate. Anion chromatography using a single-column system is a convenient and relatively inexpensive method with sufficient sensitivity for the determination of inorganic sulfate concentrations in urine, serum, and cerebrospinal fluid.     

Isolation,purification and partial characterisation of prealbumin from cerebrospinal fluid

Prealbumin from human cerebrospinal fluid was purified using a combination of ammonium sulphate precipitation, phenol precipitation, Polyacrylamide disc gel electrophoresis and gel filtration on Sephadex G-100. The homogeneity of the purified protein was established by Polyacrylamide gel electrophoresis and Immunoelectrophoresis. On the basis of its molecular weight (55,000), amino acid composition, electrophoretic mobility and immunological cross-reactivity, the prealbumin from cerebrospinal fluid showed complete identity with serum prealbumin. The cerebrospinal fluid prealbumin levels in various neurological disorders may have a diagnostic significance. Part of the Ph. D. thesis submitted by the first author.     

Brain changes in BDNF and S100B induced by ketogenic diets in Wistar rats

AimsWe investigated the effects of ketogenic diet (KD) on levels of tumor necrosis factor alpha (TNF-α, a classical pro-inflammatory cytokine), BDNF (brain-derived neurotrophic factor, commonly associated with synaptic plasticity), and S100B, an astrocyte neurotrophic cytokine involved in metabolism regulation.Main methodsYoung Wistar rats were fed during 8 weeks with control diet or two KD, containing different proportions of omega 6 and omega 3 polyunsaturated fatty acids. Contents of TNF-α, BDNF and S100B were measured by ELISA in two brain regions (hippocampus and striatum) as well as blood serum and cerebrospinal fluid.Key findingsOur data suggest that KD was able to reduce the levels of BDNF in the striatum (but not in hippocampus) and S100B in the cerebrospinal fluid of rats. These alterations were not affected by the proportion of polyunsaturated fatty acids offered. No changes in S100B content were observed in serum or analyzed brain regions. Basal TNF-α content was not affected by KD.SignificanceThese findings reinforce the importance of this diet as an inductor of alterations in the brain, and such changes might contribute to the understanding of the effects (and side effects) of KD in brain disorders.     

Effects of cerebrospinal fluid preparation from male and female patients with opiate dependence on rat open-field behavior

The effects of the intranasal administration of preparations made from the cerebrospinal fluid of male and female opiate users on the open-field rat behavior were studied. Behavioral differences were demonstrated in the effects of preparations from female and male cerebrospinal fluid. The administration of the "male" preparation produced a significant decrease in the locomotor activity and increase in the immobilization time and grooming duration, while the "female" preparation had the opposite effects. These differences may result from different content of endogenous and exogenous opiates and dopamine (and its metabolites) in the cerebrospinal fluid of male and female opiate users.     

Use of a novel double-sandwich enzyme-linked immunosorbent assay method for assaying chondroitin sulfate proteoglycans that bear 3-nitrotyrosine core protein modifications, a previously unrecognized proteoglycan modification in hydrocephalus

3-Nitrotyrosine is a useful marker for nitric oxide-mediated tissue injury. However, which proteins are preferred peroxynitrite modification targets is unclear. Chondroitin sulfate proteoglycans (CSPGs) abnormally accumulate in cerebrospinal fluid of human neonates with hydrocephalus and may be a target for peroxynitrite modification. We examined (1). whether CSPG core protein can be modified by peroxynitrite in vitro; (2). to what degree in comparison to bovine serum albumin (BSA), the most commonly used nitrated protein standard; (3). whether nitrated CSPGs can be measured directly in biological samples; and (4). whether nitrated proteoglycan concentrations in cerebrospinal fluid correlate with disease. In vitro nitration of bovine aggrecan was performed by exposure to different peroxynitrite concentrations, and 3-nitrotyrosine products were measured. Bovine serum albumin (BSA) nitration was also performed in comparison. A larger percentage of tyrosine residues were nitrated in aggrecan than in BSA under all conditions tested. An enzyme-linked immunosorbent assay (ELISA) for 3-nitrotyrosine consistently overestimated aggrecan nitration when nitrated BSA was used as the standard. This is important as most current assays of nitration in biological samples use nitrated BSA as the standard. Therefore, if nitrated CPSGs were a substantial portion of the nitrated proteins in a sample, total nitrated protein content would be overestimated. Aggrecan retained its function of binding hyaluronic acid despite substantial nitration. A double-sandwich ELISA was developed for nitrated CSPGs in biological samples, using nitrated aggrecan as standard. [Nitrated CSPG] was found to be significantly elevated in preterm hydrocephalus cerebrospinal fluid (P<0.02), but correlated poorly with cerebrospinal fluid [nitric oxide] (P>0.069), suggesting that nitrated CSPG and NO levels may be independant markers of tissue injury. Peroxynitrite-mediated protein tyrosine nitration is a previously unrecognized modification of CSPGs, and may reflect level of brain injury in hydrocephalus.