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Gene transfer into mammalian central nervous system using herpes virus vectors: extended expression of bacterial lacZ in neurons using the neuron-specific enolase promoter.

A herpes simplex virus (HSV) vector in which the mammalian promoter for neuron-specific enolase (NSE) controls expression of a marker gene was analyzed for its ability to drive expression of this foreign gene in culture and in vivo. In cultured cells, the vector appeared to give neuron-specific expression. Introduction of 10(6) pfu of the virus into the adult rat caudate nucleus by stereotactic injection was not toxic to the animals and yielded beta-galactosidase (beta-gal)-positive neurons for at least 30 days after viral inoculation. This recombinant herpes virus vector is the first described to use a mammalian promoter to yield extended expression of a foreign gene product in the adult mammalian central nervous system (CNS).

Animals↗

Neuron-specific enolase expression and neuroendocrine differentiation in carcinomas of the breast.

The concentration of neuron-specific enolase (NSE, gamma-subunit of enolase) in tumor extracts of 25 nonargyrophilic and seven argyrophilic carcinomas of the female breast was measured by radioimmunoassay. The mean concentration of NSE in nonargyrophilic carcinomas was 89 ng/mg of protein, (range, 9 to 281 ng/mg), and the mean concentration of NSE in argyrophilic carcinomas was 153 ng/mg of protein (range, 4 to 420 ng/mg). All carcinomas, irrespective of the silver reaction, could be grouped in regular histopathologic terms. Neuron-specific enolase immunoreactivity in sections of the corresponding fixed tumors occurred in both silver-stained and unreactive tumors, although the reaction was more prominent in the argyrophilic tumors. The results indicate that NSE in considerable amounts may be present in both argyrophilic and nonargyrophilic carcinomas of the breast, although higher concentrations are found most often in the former tumor type.

Aged↗

The neural marker neuron-specific enolase in the development of embryo chicken retina: a morphological description.

The localisation and distribution of Neuron-specific enolase is reported in the avian Gallus domesticus retinal development by using immunocytochemistry. Neuron specific enolase was found to be present from the early days of incubation to the post-hatch period. The results obtained using this neural marker showed the development pattern of the distribution and the sequence of differentiation of the retinal neural structures. Like the finding of the members of the phylogenetic scale, this enzyme should prove to be a useful tool in the neural development of the chicken retina.

Animals↗

[Gastrin-releasing peptide as a specific tumor marker in patients with small cell lung carcinoma, compared with neuron-specific enolase].

A sensitive radioimmunoassay (RIA) system for plasma gastrin-releasing peptide (GRP) was developed using immune-affinity chromatography for plasma extraction. Plasma neuron-specific enolase (NSE) levels were determined by use of RIA without extraction. Plasma GRP levels in 12 control subjects were (mean +/- SD) 1.2 +/- 0.26pg/ml. Plasma GRP levels were elevated at frequencies of 79% in untreated patients with small cell lung carcinoma (SCLC). The levels in 21 patients with non-SCLC were not elevated. In nine of 10 patients with complete response (CR) or partial response (PR), plasma GRP levels decreased significantly when the patients were judged to have achieved CR or PR. In four patients with progressive disease (PD), the levels were elevated after treatment when compared with levels before treatment. In six of 10 patients with CR or PR, plasma NSE levels decreased significantly at the judgment of CR or PR. In two of four patients with PD, the levels were elevated after treatment. Furthermore, changes in plasma GRP level showed more excellent correlation with the therapeutic response than changes in plasma NSE level in the clinical courses of two patients with CR and a patient with PD. These results suggest that the plasma GRP level could be a useful tumor marker in SCLC patients.

Biomarkers, Tumor↗

Neuron-specific enolase in neuroblastoma and other pediatric tumors: a comparative nude mouse and clinical investigation.

Neuron-specific enolase (gamma-subunit isozyme) in extracts of human neuroblastomas and other tumors in children was measured by means of a radio-immunoassay in order to elucidate the rationale for clinical determination of its serum concentrations in patients. Surgical specimens as well as tumors xenotransplanted in nude mice were studied. The neuron-specific enolase (NSE) concentrations in extracts of xenografted and surgical neuroblastomas were 1,733.2 +/- 677.3 (range; 883-2,659) ng/mg protein and 1,735.3 +/- 941.1 (559-2,933) ng/mg protein, respectively, while other pediatric tumors examined showed much lower levels of NSE: 121.5 +/- 146.1 (22-374) ng/mg protein in xenografts and 17.7-610 ng/mg protein in surgical specimens. Sera of nude mice bearing human neuroblastomas also showed NSE concentrations ranging from 1,102 to 4,400 ng/ml, very much higher than those in sera of nude mice without tumors or with other pediatric tumors. These results tend to confirm the specificity of NSE for neuroblastoma, and the difference in NSE concentrations among five neuroblastoma xenografts is discussed in relation to their cytogenetic characteristics.

Animals↗

Neuron-specific enolase in the pituitary gland.

Neuron-specific enolase (NSE) is present in many types of peptide-secreting neuroendocrine cells and in tumours derived from them, but little work has been done on the pituitary gland. Serial sections of normal rat (n = 9) and human (n = 7) pituitary gland, spontaneous rat pituitary tumours (n = 14) and human pituitary tumours, both hormonally active (n = 7) and inactive (n = 10), were immunostained for NSE and the 6 major anterior pituitary hormones. The neural lobe stained strongly and the intermediate lobe variably for NSE. In the anterior lobe, NSE immunoreactivity was present with variable intensity in the majority of hormone-producing cells of all six types. Cells with strong hormone immunoreactivity were usually only moderately stained for NSE. All the human and rat pituitary adenomas examined were positively stained for NSE, though to varying degrees. The pituitary gland is thus no exception to the rule that NSE is found in peptide-secreting neuroendocrine cells and their tumours.

Acromegaly↗

Neuronal-specific roles of the survival motor neuron protein: evidence from survival motor neuron expression patterns in the developing human central nervous system.

Despite recent data on the cellular function of the survival motor neuron (SMN) gene, the spinal muscular atrophy (SMA) disease gene, the role of the SMN protein in motor neurons and hence in the pathogenesis of SMA is still unclear. The spatial and temporal expression of SMN in neurons, particularly during development, could help in verifying the hypotheses on the SMN protein functions so far proposed. We have therefore investigated the expression and subcellular localization of the SMN protein in the human central nervous system (CNS) during ontogenesis with immunocytochemical, confocal immunofluorescence, and Western blot experiments using a panel of anti-SMN antibodies recognizing the full-length SMN protein. The experiments not only revealed the early SMN expression in all neurons, but also demonstrated the progressive shift in SMN subcellular localization from mainly nuclear to cytoplasmic and then to axons during CNS maturation. This finding was present in selected neuronal cell populations and it was particularly conspicuous in motor neurons. Our data support the idea of a specific role for SMN in axons, which becomes predominant in the ontogenetic period encompassing axonogenesis and axonal sprouting. In addition, the asymmetric SMN staining demonstrated in the germinative neuroepithelium suggests a possible role for SMN in neuronal migration and/or differentiation.

Adult↗

Clinical relevance of increased neuron-specific enolase concentration in cerebrospinal fluid.

Neuron-specific enolase (NSE) concentrations in cerebrospinal fluid (CSF) and serum have been studied by an EIA-method using monoclonal antibodies against human NSE. In a control group (n = 24) the mean NSE value (+/- SD) in CSF was 10.8 (+/- 4.5) ng/ml. Increased NSE values in CSF (greater than or equal to 20 ng/ml, ie greater than or equal to means + 2s) have been detected in 33/172 patients with the following neurological diseases: CNS tumors (6/30), infarctions (6/36), cerebral ischemias (5/25), inflammatory diseases (7/33), epilepsias (3/10) and miscellaneous neurological diseases (6/38). The NSE assay in CSF was not specific for a single neurological disease. In 9% of all patients with an organic neurological disease the increased NSE concentration was the only abnormal result in the CNS out of variables routinely determined in CSF. The discrimination between an organic and psychogenic origin of epilepsy may be possible by an NSE analysis in CSF. The NSE assay in CSF can be recommended as an unspecific screening parameter for pathological organic CNS processes.

Antibodies, Monoclonal↗

Neuron-specific enolase.

The clinical value of neuron-specific enolase as a marker is small cell lung cancer, neuroblastoma, melanoma and seminoma has been reviewed. The role of serum and cerebrospinal NSE in benign and malignant disease of the central nervous system is discussed.

Biomarkers↗

Serum neuron-specific enolase in human status epilepticus.

Neuron-specific enolase (NSE) is a sensitive marker of brain injury after stroke, global ischemia, and coma. We report changes in serum NSE (s-NSE) in 19 patients who sustained status epilepticus. s-NSE peaked within 24 to 48 hours after status epilepticus. The mean peak s-NSE level for the entire group was elevated compared with the levels for normal controls (24.87 ng/ml versus 5.36 ng/ml, p = 0.0001) and for epileptic controls (24.87 ng/ml versus 4.61 ng/ml, p = 0.0001). The mean peak s-NSE level for the 11 subjects without an acute neurologic insult (15.44 ng/ml) was also significantly increased compared with levels for normal and epileptic controls. Further, s-NSE was significantly correlated with outcome and duration. We conclude that s-NSE is a promising in vivo marker of brain injury in status epilepticus and warrants further study in larger populations.

Humans↗

Human neuron-specific enolase: genetic and developmental studies.

Neuron-specific enolase is readily detected as early as 54 days gestational age in brain extracts from human fetuses and undergoes a rapid increase with developmental maturation of the nervous system. No genetic variants of this enzyme were found among 14 fetal and 120 postnatal specimens, consistent with previous findings of highly restricted variation among glycolytic enzymes.

Cerebral Cortex↗

[Determination of various tumor markers, with special reference to neuron-specific enolase in lung cancer].

Serum neuron-specific enolase (NSE) was measured in 23 patients with small cell lung cancer (SCLC) and 184 patients with non-small cell lung cancer (non-SCLC), both of which were untreated. Increased levels of serum NSE were observed in 82.6% (19/23) of SCLC, whereas 9.8% (18/184) of non-SCLC had positive results showing an overall positive rate of 17.9% (37/207) in lung cancer cases. In addition, the elevation of serum NSE levels in non-SCLC patients seemed to suggest poor prognosis. Elevated serum NSE levels returned to normal with either surgical resection of the tumor or response to chemotherapy, after which serum NSE levels were again raised to levels higher than the previous ones in cases of relapse or progression. The evaluation of serum NSE may be a useful marker for both diagnosis and monitoring of responsiveness to therapy as well as for recognition of relapse and progression in SCLC. Identification of NSE as assessed by immunohistochemical procedure employing the ABC method on formalin-fixed paraffin-embedded tissue sections in lung cancer cases of each histological type, showed that some materials from non-SCLC cases were positively stained despite the presence of normal serum NSE levels, and did not always parallel the serum levels. Among other various tumor markers determined, serum CA 19-9 had a relatively high positive rate of 38.2% (42/110) in adenocarcinoma of the lung.

Antigens, Neoplasm↗

[Usefulness of neuron specific enolase in patients with head and neck malignant tumors].

The authors present results of evaluation of neuron specific enolase (NSE) in 146 persons. We analyzed a concentration of NSE in preliminary study and in monitoring depending on clinical advances of tumour, mass of primary tumour and metastases to lymph nodes. Neuron specific enolase was excluded because the number of positive results was the same both in the main and control group. Our results indicate that evaluation of NSE have not height value in monitoring patients with head and neck carcinoma.

Adolescent↗

Neuron-specific enolase and Leu-7 immunoreactive small round-cell neoplasm. The relationship to Ewing's sarcoma in bone and soft tissue.

The authors present an immunohistochemical analysis of tissue from five cases of morphologically distinctive Ewing's sarcoma in bone and soft tissue. The mean age of the five patients was 16.6 years, with a range of 6-28 years. The tumors existed in chest wall, pelvis, and lower extremities. Two siblings with tumor in bone were clinically diagnosed as having Ewing's sarcoma. All cases had immunoreactivity for neuron-specific enolase (NSE), and four cases revealed positive staining for Leu-7. Neuron-specific enolase is highly specific for neurons and neuroendocrine cells. In addition, immunoreactivity for Leu-7, expressing a natural killer activity, has been demonstrated in peripheral nerve fibers and neuroendocrine cells. The authors suggest that NSE and Leu-7 immunoreactive small round-cell neoplasm is probably a primitive neuroectodermal tumor and should be categorized as Ewing's sarcoma in bone and soft tissue.

Adolescent↗

Cerebrospinal fluid and serum neuron-specific enolase levels after febrile seizures.

PURPOSE: Neuron-specific enolase (NSE) has been established as a reliable marker of neuronal damage in various neurologic disorders. The aim of this study was to evaluate whether febrile seizures (FS) cause brain damage, based on the serum and cerebrospinal fluid (CSF) levels of NSE. METHODS: Fifty-three patients aged from 6 months to 7 years were enrolled. Among them, 36 patients had generalized seizures, and 17 had partial seizures. The maximal seizure duration was 90 min. Blood and CSF samples for measurement of NSE were obtained immediately after the seizure. NSE was measured using an enzyme immunoassay (EIA). RESULTS: Serum and CSF levels of NSE ranged up to 10 ng/mL, but very high levels were not observed. In patients with partial seizures, the NSE level in the CSF and the ratio of the CSF to serum NSE levels showed a strong correlation with seizure duration. Conversely, there were no correlations between NSE levels and seizure duration in the patients with generalized seizures. CONCLUSIONS: These results indicate that FS seldom cause severe neurologic damage, but prolonged partial seizures may cause slight neuronal injury.

Biomarkers↗

The effect of a neuron-specific antiserum, BPM, on the in vitro development of cerebellar granule cells.

Anti-BPM is a neuron-specific antiserum which specifically recognizes the D2 cell adhesion molecule in crossed immunoelectrophoresis of Triton X-100-solubilized brain extracts. Here the effect of this antiserum on the in vitro development of cerebellar neuronal cultures is described. The initial adhesion of cells and neurite outgrowth were not influenced by immunoglobulin fractions of anti-BPM. However, after 5 days in vitro the cultures had become completely disorganized, with the majority of cells being dead at immunoglobulin concentrations greater than 0.5 mg/ml culture medium. This effect was seen only with immunoglobulins and their F(ab')2 fragments, the F(ab') fragments being without effect. The addition of anti-BPM to 8-day-old cultures resulted in a more rapid and pronounced rate of cell death. In many instances this was preceded by a rapid "destabilization" of culture organization. The cytotoxic effect of anti-BPM was neuron specific and the small numbers of astrocytes and fibroblasts found in the cultures were unaffected by prolonged exposure to this serum.

Animals↗

Neuron-specific enolase: investigation on its possible retrograde axonal transport.

The possible rapid retrograde axonal transport of neuron-specific enolase was investigated employing a sensitive radioimmunoassay. The right sciatic nerve of rats was ligated and 6 and 24 h later the nerve was cut in two 5.0 mm segments immediately proximal and distal to the ligature. Similar segments were taken from the contralateral nerve as a control. No increase in the amount of neuron-specific enolase in the right-distal segment was observed indicating that the protein is not a component of retrograde axonal transport.

Animals↗

Immunoradiometric assay for alpha gamma- and gamma gamma-enolase (neuron-specific enolase), with use of monoclonal antibodies and magnetizable polymer particles.

Monoclonal antibodies were raised against neuron-specific enolase, gamma gamma-enolase, and used in an immunoradiometric assay (IRMA), with mono-disperse magnetizable particles as the solid phase. The assay's sensitivity was 0.4 microgram/L and the interassay coefficient of variation was less than 5% in the working range from 0.4 to 170 micrograms/L. Compared with our radioimmunoassay based on polyclonal antibodies, the incubation time is shorter, and precision and sensitivity are improved. The IRMA also improved detection of neuron-specific enolase in sera from patients with lung cancer without a concomitant change in measured enolase in the reference population. The better sensitivity of the IRMA results from its ability to measure alpha gamma- and gamma gamma-enolase with equal response. Ninety percent of the small-cell lung carcinoma patients (36 of 40) had increased values before treatment, compared with 7% of non-small-cell lung carcinoma patients (8 of 114).

Animals↗