Interference by platelets in the electrophoretic determination of enolase isoenzymes in plasma

Serum and plasma gave different electrophoretic patterns when enolase isoenzymes were evaluated by electrophoresis on cellulose acetate. Plasma isoenzyme bands were more intense, and there was an additional one (band P) that was not present in serum. We show that, under the conditions of electrophoresis, some of the residual platelets in the plasma are ruptured, releasing intracellular enolases and consequently leading to intensification of the isoenzyme bands. The band P originated from the remaining unruptured platelets. Thus plasma samples must be platelet-free for determination of enolase isoenzyme to be reliable.     

Clinical value of an amplified electrophoretic determination of enolase isoenzymes in small cell lung carcinoma patients

Neurone specific enolase (NSE); alpha gamma and gamma gamma isoenzymes of the glycolytic enzyme enolase, is found in considerable quantity in serum of patients with small cell lung cancer (SCLC). The spectrophotometric measurement of serum enolase activity, plus the electrophoretic separation of isoenzymes (alpha alpha, alpha gamma and gamma gamma) using an amplification reaction constitute a simple method, the application of which has not yet been demonstrated. In patients, serum values of higher than 25 U/l of total enolase activity, with more than 10% NSE, were considered to be positive. Seventy patients diagnosed as SCLC were classified before treatment as having either limited (LD) or extensive (ED) disease, and after chemotherapy as being in complete (CR) or partial remission (PR), stable state (SS) or in relapse (R). The levels of enolase activity and NSE (M +/- SE) in these patients (enolase: 67 +/- 7 U/l, NSE 27 +/- 2%) were different from those in a control group of 19 patients with non-small cell lung cancer (enolase: 29 +/- 2 U/l, NSE: 7 +/- 1%) (p less than 0.001) at the time of diagnosis. Mean enolase and NSE levels in patients with SCLC were seen to differ significantly according to the clinical stage. The results of those patients with ED differed from those of patients with LD (p less than 0.001). The results of the group of patients that achieved remission differed from that of patients during relapse (p less than 0.0001). Serial measurements demonstrated a good correlation between enolase and NSE serum levels and the progression of the disease. The usefulness of this method in the early assessment of treatment was also demonstrated. The clinical usefulness of the dosage of NSE with that of two other tumour markers CKBB and mitochondrial CK was compared.     

Cerebrospinal fluid and serum neuron-specific enolase in acute benign headache

We determined the cerebrospinal fluid (CSF) and serum neuron-specific enolase (NSE) concentrations in 19 patients with acute benign headache. All patients had normal neurological examination, CSF and head computed tomography scan. The final diagnoses were: primary thunderclap headache ( n  = 7), primary exertional headache ( n  = 3), primary cough headache ( n  = 1), migraine without aura ( n  = 4), headache unspecified ( n  = 2), probable infrequent episodic tension-type headache ( n  = 1), headache attributed to hypertensive crisis without hypertensive encephalopathy ( n  = 1). A group of 108 healthy subjects served as controls. CSF NSE concentration was 14.16 ng/ml [95% confidence interval (CI) 11.86, 16.47)] in the headache sample (controls 17.19 ng/ml, 95% CI 16.23, 18.15). Serum NSE concentration was 7.50 ng/ml (95% CI 5.20, 9.80) in the headache sample (controls 8.45 ng/ml, 95% CI 7.67, 9.23). CSF/serum ratio was 2.81 (95% CI 2.21, 3.40) in the headache sample (controls 2.23, 95% CI 2.03, 2.42). Acute benign headache is not associated with neuronal damage as estimated by means of CSF and serum NSE concentration.     

Sensitive, simple staining method for use in electrophoretic determination of amylase isoenzymes in serum

We have developed a sensitive and simple staining method for use in electrophoretic analysis of serum alpha-amylase (EC 3.2.1.1) isoenzymes. The principle of this method is as follows. alpha-Amylase hydrolyzes 4-nitrophenylmaltoheptaoside to generate oligosaccharide, which is then converted to gluconolactone in the presence of oligosaccharide dehydrogenase (no EC no. assigned), with concomitant reduction of 1-methoxy-5-methylphenazinium methylsulfate (1-m-PMS) to 1-m-PMSH. The hydrogen in 1-m-PMSH is then transferred to 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide to yield formazan. Each of the isoenzymes can then be measured densitometrically. The mean (and SD) values for total amylase, P1, S1, and S2 as determined by this method with sera from 25 healthy adults in fasting were 251 (64), 104 (35), 126 (40), and 22 (11) U/L, respectively. Between-assay CVs (n = 10) for determinations of P1, S1, and S2 were 3.54%, 4.03%, and 7.01%, respectively.     

Clinical roles of serum autoantibody against neuron-specific enolase in glaucoma patients

In our recent papers, we found the presence of serum autoantibody against neuron specific enolase (NSE) in some glaucoma patients and suggested that this antibody might have significant roles in pathogenesis of glaucomatous optic neuropathy. In order to evaluate further the clinical roles of serum autoantibody against NSE in glaucoma, serum autoantibody against NSE was examined by western blot analysis and enzyme-linked immunosorbent assay (ELISA) in 4 patients with ocular hypertension (OH) and 242 patients with glaucoma (normal tension glaucoma [NTG], 73 cases; primary open angle glaucoma [POAG], 169 cases), and the relationships between the titers of anti-NSE antibody and clinical characteristics were evaluated. The titers of anti-NSE antibody showed a regular decreasing pattern with deteriorating visual field losses and glaucoma stages in POAG, especially early and late stages. However, no systematic pattern was observed in NTG. Although maximum and mean intraocular pressures (IOP)s and progression of visual field losses showed no correlation with the levels of serum anti-NSE antibody titers in either POAG or NTG, the anti-NSE antibody titers were relatively higher in NTG with visual field deterioration than in those without it. The present observations suggest that serum autoantibody against NSE may be clinically useful for diagnosing early stages of POAG, and for monitoring glaucoma progression of NTG.     

血清神经特异性烯醇化酶与脑胶质瘤患者术后脑损伤的相关性

目的探讨脑胶质瘤患者手术前、后血清神经特异性烯醇化酶(neuron-specific enolase, NSE)水平与脑组织损伤的关系。方法 48例经手术治疗的脑胶质瘤患者,于术前1 d及术后1、3、7 d分别检测血清NSE水平,并进行手术前、后比较;收集手术前、后影像学资料,采用Pearson或Spearman相关分析血清NSE水平与肿瘤体积、脑组织水肿体积及术后6个月KPS功能评分的相关性。结果术后1、3、7 d时患者血清NSE水平[(15.07±2.34)、(18.20±4.31)、(15.20±3.94)μg/L]均高于术前[(12.98±2.61)μg/L](P<0.05),术后3 d时高于术后1 d时(P<0.05),术后7 d与术后1 d比较差异无统计学意义(P>0.05);不同部位、不同级别脑胶质瘤患者手术前、后血清NSE水平比较差异均无统计学意义(P>0.05);术前血清NSE水平与瘤周水肿体积呈正相关(r=0.757,P<0.001),与肿瘤体积无相关性(r=0.190,P=0.195);术后3 d血清NSE水平与术后3 d脑水肿体积呈正相关(r=0.553,P=0.002),术后7 d血清NSE水平与术后6个月KPS评分(80分)呈负相关性(r=-0.821,P<0.001)。结论血清NSE水平可辅助评估脑胶质瘤手术患者的脑损伤程度变化。     

Improved agarose electrophoretic method for separating alkaline phosphatase isoenzymes in serum

A modified agarose electrophoretic system for the separation of alkaline phosphatase (ALP, EC 3.1.3.1) isoenzymes is described. Bone, liver, high-molecular-mass, and intestinal ALP are separated with high reproducibility. The sensitivity of the agarose system is superior to cellulose acetate in detecting high-Mr ALP. Correlation is good between bone ALP fractions scanned before and after treatment with neuraminidase. Immunoglobulin-bound ALPs, the ALP-lipoprotein-X complex, and the additional ALP fraction observed in transient hyperphosphatasemia in children are detected by their peculiar electrophoretic mobility in the proposed system. Approximately 25% of the samples contained an additional fraction of intestinal-type ALP, as evidenced by neuraminidase treatment and use of polyclonal and monoclonal antibodies. Because the electrophoretic mobilities of this "intestinal variant" and of some immunoglobulin-bound ALP fractions are identical to those of bone and intestinal ALP, respectively, treatment of the samples with a polyclonal antibody that reacts with intestinal ALP is advised.     

保存温度、时间、溶血对血清标本中NSE测定的影响

目的探讨血清标本保存时间、保存温度和溶血对神经元特异性烯醇化酶(NSE)测定的影响。方法将正常人血清标本分别置于4℃和-20℃,用电化学发光(双抗体夹心)法检测放置当天、第3天、第5天的NSE水平,比较不同保存温度、保存时间条件下,非溶血血清与溶血血清中NSE值的变化情况。结果在相同温度下,放置第3天的NSE检测结果高于当天的检测结果(t分别=2.99、6.90,P均<0.05);而放置第5天检测结果较放置第3天的检测结果进一步升高(t分别=1.97、2.04,P均<0.05)。在相同放置时间下,只有在放置第5天,4℃组的NSE检测结果高于-20℃组,差异具有统计学意义(t=4.67,P<0.05)。不同保存时间、不同保存温度及两者交互效应对于NSE检测结果均有明显影响(P均<0.05)。在相同保存温度、相同放置时间下,当天-20℃、放置第3天4℃和-20℃、放置第5天4℃和-20℃的溶血标本和非溶血标本NSE水平比较,差异均有统计学意义(t分别=2.16、3.49、2.60、13.66、8.42,P均<0.05)。结论血清NSE含量会因溶血、全血标本放置时间的延长以及保存温度的升高而增高。     

短暂性脑缺血发作患者血清神经元特异性烯醇酶水平的动态变化

背景：神经元特异性烯醇酶是γ型同工酶特异地存在于神经元和神经内分泌细胞的胞浆内，是神经元损伤较敏感的标志物。目的：观察短暂性脑缺血发作患者血清神经元特异性烯醇酶的变化，探讨其与疾病的神经损伤程度的关系。设计：病例一对照观察。单位：济南市第四人民医院神经内科。对象：选择2002-03／2004—05在济南市第四人民医院神经内科入院的短暂性脑缺血发作患者29例（均为起病后6h的急诊观察患者）。男18例，女11例。平均（60．36&;#177;11．67）岁。根据神经功能缺失症状持续时间分为两组：短症状组（≤6h）19例，长症状组（〉6h）10例。同期选取健康体检者为对照组，共25人。男15人，女10人。平均（62．34&;#177;9．65）岁。方法：对照组只取1次空腹肘静脉血1mL，短暂性脑缺血组在入院即时，第2，3，4，5天分别采空腹肘静脉血1mL。采用罗氏Elecsys2010免疫测定分析仪检测血清神经元特异性烯醇酶。用神经功能缺损程度量表（基本痊愈为评分减少90％-100％；显著进步为评分减少46％-89％；进步为评分减少18％-45％；无效为评分减少17％以下或病情恶化）评定患者的神经损伤程度。主要观察指标：观察患者血清神经元特异性烯醇酶的每天变化情况。结果：检测者54例均进入结果分析。①神经元特异性烯醇酶浓度的比较：短暂性脑缺血发作组明显高于对照组[（23．53&;#177;12．35，14．29&;#177;6．83）μg／L，t=2．678，P〈0．01]。②急性期内神经元特异性烯醇酶变化曲线：早期增高，次日达高峰，后趋向逐渐下降，四五天基本恢复正常。③神经功能缺失症状持续时间不同的两组血清神经元特异性烯醇酶：短症状组明显高于对照组[（19．24&;#177;8．95，14．29&;#177;6．83）μg／L，t=1．893，P〈0．05]，长症状组高于对照组[（28．87&;#177;13．15，14．29&;#177;6.83）μg／L，t=4．367，P〈0．001]。④神经元特异性烯醇酶值的大小与神经功能缺损时间呈正相关（r=0．815，P〈0．01）。结论：短暂性脑缺血发作患者短期内血清神经元特异性烯醇增高，24～36h达到高峰，提示检测短暂性脑缺血发作患者血清神经元特异性烯醇酶浓度。对判断病情严重程度有指导意义。     

短暂性脑缺血发作患者血清神经元特异性烯醇酶水平的动态变化

背景神经元特异性烯醇酶是γ型同工酶特异地存在于神经元和神经内分泌细胞的胞浆内,是神经元损伤较敏感的标志物.目的观察短暂性脑缺血发作患者血清神经元特异性烯醇酶的变化,探讨其与疾病的神经损伤程度的关系.设计病例-对照观察.单位济南市第四人民医院神经内科.对象选择2002-03/2004-05在济南市第四人民医院神经内科入院的短暂性脑缺血发作患者29例(均为起病后6 h的急诊观察患者).男18例,女11例.平均(60.36±11.67)岁.根据神经功能缺失症状持续时间分为两组短症状组(≤6h)19例,长症状组(＞6h)10例.同期选取健康体检者为对照组,共25人.男15人,女10人.平均(62.34±9.65)岁.方法对照组只取1次空腹肘静脉血1 mL,短暂性脑缺血组在入院即时,第2,3,4,5天分别采空腹肘静脉血1 mL.采用罗氏Elecsys2010免疫测定分析仪检测血清神经元特异性烯醇酶.用神经功能缺损程度量表(基本痊愈为评分减少90%-100%;显著进步为评分减少46%-89%;进步为评分减少18%-45%;无效为评分减少17%以下或病情恶化)评定患者的神经损伤程度.主要观察指标观察患者血清神经元特异性烯醇酶的每天变化情况.结果检测者54例均进入结果分析.[1]神经元特异性烯醇酶浓度的比较短暂性脑缺血发作组明显高于对照组[(23.53±12.35,14.29±6.83)μg/L,t=2.678,P＜0.01].[2]急性期内神经元特异性烯醇酶变化曲线早期增高,次日达高峰,后趋向逐渐下降,四五天基本恢复正常.[3]神经功能缺失症状持续时间不同的两组血清神经元特异性烯醇酶短症状组明显高于对照组[(19.24±8.95,14.29±6.83)μg/L,t=1.893,P＜0.05],长症状组高于对照组[(28.87±13.15,14.29±6.83)μg/L,t=4.367,P＜0.001].[4]神经元特异性烯醇酶值的大小与神经功能缺损时间呈正相关(r=0.815,P＜0.01).结论短暂性脑缺血发作患者短期内血清神经元特异性烯醇增高,24～36h达到高峰,提示检测短暂性脑缺血发作患者血清神经元特异性烯醇酶浓度.对判断病情严重程度有指导意义.     

Cerebrospinal fluid neuron-specific enolase in non-selected patients

The degree to which cerebrospinal fluid (CSF) neuron-specific enolase (NSE) contributes to the diagnosis and prognosis of disorders of the central nervous system (CNS) or peripheral nervous system (PNS) is still under debate. The aim of the study was thus to assess the validity of CSF-NSE levels in the diagnostic work-up of these conditions. The study consecutively included 106 adult patients who had undergone a diagnostic spinal tap or myelography during the diagnostic work-up for various CNS or PNS disorders. Thirty-five of these patients (16 F, 19 M, aged 24-88 years) without indication of a CNS disorder and with normal routine CSF investigations served as controls. The remaining 71 patients (31 F, 40 M, aged 28-87 years) constituted the disease group. CSF-NSE was independent of sex and age. The upper reference limit of CSF-NSE was 0.01536 ng/L. CSF-NSE was elevated in 13 of the 71 patients (18%): 6 with metabolic myopathy, 4 with polyneuropathy and 3 with hepatic encephalopathy, multiple sclerosis and paraspasticity, respectively. Only 6 of the 13 patients (46%) showed CNS involvement. The study shows that CSF-NSE is elevated in only one-fifth of unselected patients who consecutively undergo a spinal tap. CSF-NSE is elevated most frequently in patients with metabolic myopathy and polyneuropathy, even in cases without CNS abnormalities.     

糖尿病周围神经病变患者检测血清神经元特异性烯醇化酶的临床价值

目的探讨糖尿病周围神经病变(DPN)患者检测血清神经元特异性烯醇化酶(NSE)的临床价值。方法采用化学发光法检测120例糖尿病并发周围神经病变患者(DPN组)、47例糖尿病未并发周围神经病变患者(DM组)、40例健康对照者(对照组)的血清NSE,并根据多伦多临床评分系统(TCSS)评分将120例DPN患者分为轻度组(35例)、中度组(51例)、重度组(34例)。结果⑴DPN组、DM组、对照组血清NSE相比差异有统计学意义(P0.05),DPN组血清NSE显著高于DM组、对照组(P0.05),DM组与对照组血清NSE差异无统计学意义(P0.05)。⑵轻度组、中度组、重度组血清NSE相比差异有统计学意义(P0.05),重度组血清NSE显著高于轻度组、中度组(P0.05),中度组血清NSE显著高于轻度组(P0.05)。结论血清NSE与DPN密切相关,血清NSE既可以用于DPN的诊断,还可以用于DPN的病情评估。     

Elevated serum S100B protein and neuron-specific enolase levels in carbon monoxide poisoning

Objective

Carbon monoxide (CO) poisoning causes cerebral and generalized hypoxia. This study aimed to assess the possible use of serum glial marker S100B protein and neuron-specific enolase (NSE) as biochemical markers of hypoxic brain damage in acute CO poisoning.

Methods

Patients with acute CO poisoning admitted to the ED of 2 training hospitals (Ankara, Turkey) were included in this cross-sectional study. Serum levels of S100B and NSE were measured on admission. The patients were divided into 2 groups (unconscious and conscious). Twenty healthy adults were included in the study to serve as controls.

Results

A total of 70 patients poisoned by CO (mean age ± SD, 36.6 ± 16.3 years; 64.3% women) were enrolled. Although S100B concentrations were higher in patients than in the control group (P = .018), no significant difference was determined between patient and control groups with respect to NSE concentrations (P = .801). A positive correlation was noted between levels of S100B and NSE (r = 0.388; P = .001). The S100B and NSE values were higher in unconscious patients than in the control group (P = .002 and P = .013, respectively). Furthermore, S100B and NSE values were higher in unconscious vs unconscious patients (P = .047 and P = .005, respectively).

Conclusion

Elevated serum S100B and NSE levels were associated with loss of consciousness in CO poisoning in this series of patients. Serum S100B and NSE may be useful markers in the assessment of clinical status in CO poisoning.     

新生儿缺氧缺血性脑病血清神经元特异性烯醇化酶的变化及临床价值

目的通过检测新生儿缺氧缺血性脑病（HIE）治疗前、后血清神经元特异性烯醇化酶（NSE）水平,探讨其在HIE中的临床价值。方法采用电化学发光法测定90例HIE患儿治疗前、后及30名正常对照儿童血清NSE水平。将90例HIE患儿按诊断标准分为轻度、中度、重度HIE组,比较各组治疗前、后NSE水平的差异。结果 HIE患儿血清NSE水平明显高于对照组（P〈0.01）,且HIE临床分度越重,血清NSE水平升高越明显（P〈0.01）。HIE各组患儿治疗后NSE水平明显低于治疗前（P〈0.05、P〈0.01）。结论测定血中NSE能及时、有效地帮助临床对新生儿HIE的脑损伤做出早期诊断和病情轻重的判断,为治疗的监测和预后提供可靠的指导参考。     

脑梗死患者血清神经元特异性烯醇化酶的动态变化及临床意义

目的　探讨缺血性脑卒中患者急性期血清神经元特异性烯醇化酶 (NSE)的动态变化及其意义。方法　临床确诊的脑梗死患者 75例 ,测量并计算其脑梗死体积 ,采集其发病后不同时间的静脉血 ,凝固后立即 3 0 0 0r/s离心 1 5min ,取血清 ,置 - 80℃保存 ,共 2 4 8份标本 ;对照组为同年龄组健康成人 ,共 30例 ,采集标本 30份。采用NSE酶联免疫分析法测定血清中NSE浓度。数据用均数±标准差表示 ,各组间比较用方差分析和t检验及相关分析法。结果　①脑梗死组和对照组NSE浓度分别为 (7 2 5± 2 1 4 )ng/mL和 (3 97± 1 1 2 )ng/mL ,具有显著差异 (P <0 0 1 ) ;②血清NSE浓度与脑梗死体积呈正相关 ,梗死体积越大 ,NSE值越高 ,各组间比较 ,均P <0 0 5 ;③脑梗死后NSE呈现动态变化 ,发病 4h内变化不明显 ,6h后开始升高 ,2 4h后明显升高 ,2d时达高峰 ,并可持续到第 5～ 7天 ,1 4d时基本恢复到正常水平 ;④血糖水平与NSE之间有一定相关性 ,血糖水平高者 ,其NSE值亦高。⑤血清NSE水平与脑梗死患者的预后相关 ,NSE高的患者 ,其 2周时的预后较差。结论　脑梗死后患者血清NSE升高并呈现一动态变化 ,它与脑梗死的体积及患者的预后相关 ,且其变化早于影像学变化 ,可以作为脑梗死程度的依据之一 ,对确定临床治疗方案具有一     

Postsynthetic modification of human enolase isoenzymes

The original form of beta beta enolase (EC 4.2.1.11) in tissue is modified to two more electrophoretically distinct forms when incubated with human serum. The three postsynthetic forms are designated beta beta 3, beta beta 2, and beta beta 1, in order of increasing anodal mobility and increasing modification. Serum and carboxypeptidases A and B all produce identical modifications of beta beta enolase but exhibit very different pH-activity profiles. A purified human serum protein previously named "modifying protein," which is responsible for the modification of creatine kinase-M and alpha-enolase subunits, modifies beta beta enolase and also has a pH-activity profile identical to that for serum. Thus we conclude that the modifying protein is not identical to either carboxypeptidase A or B; it may, however, be an as-yet-undescribed carboxypeptidase. With increased modification, both alpha alpha and beta beta enolase decrease in apparent activation energy; gamma gamma enolase shows no evidence of modification, and its apparent activation energy remains stable. Measurement of activation energy is an easy tool for screening for postsynthetic modifications in an enzyme.