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PubMed1999

Glial heterotopia in the subcutaneous tissue overlying T-12.

Heterotopic glial nodules occur most commonly in the head and neck area, and are theorized to arise following abnormalities in the development of the facial and skull bone plates. However, in spite of the fact that some of these lesions are associated with communication with the central nervous system (CNS), the lack of a meningeal component, argues against simple herniation and separation of brain tissue through a defect in the skull. We present an infant with a nodule directly over the spine present in the T-12 region with no skin abnormalities. Magnetic resonance imaging (MRI) and computerized axial tomography (CT) showed no spinal abnormalities with an overlying fibrotic soft tissue mass. The patient had no other associated clinical findings. Histologic findings showed a cellular component arising within the reticular dermis with a deep circumscribed margin. The nodule contained irregularly shaped cells containing abundant cytoplasm and indistinct cellular margins with bland nuclei. These cells were clustered around and between a fibro-mucinous stroma. Immunohistochemical stains showed positive staining for S-100 protein, vimentin, GFAP, NSE, and CD57, and negative staining for Ki-67, CD34, Neurofilament protein, cytokeratin, and EMA. The spindle cells showed positive staining for CD34 and vimentin. The clinical and histologic features and immunohistochemical profiles are used to separate this lesion from the closely related, ependymal rests, ependymomas, and primary cutaneous chorodomas.

CD57 Antigens↗

PubMedDate not supplied

[Brain emboli in the lungs of cattle].

There is no information whether the BSE agent is introduced into the human food chain through contamination of the lungs of cattle with central nervous system tissue (CNS). Studies in the United Kingdom and in the USA showed that CNS tissue could contaminate the lungs after using pneumatic powered air injection stunners (e.g. "The Knocker") or after pithing. Thus, pithing was forbidden in the European Union since January 2001. In German abattoirs conventional cartridge-fired stunners (e.g. model by Schermer) are usually applied. Pithing was used up to December 2000 in approx. 75% of the German abattoirs. In the present study 323 lungs of cattle were analysed for CNS. The lungs were derived from cattle exclusive stunned by use of the knocker from Schermer. 60% of the lungs contained emboli which were tested with immuno chemistry as well as immuno histochemistry to detect CNS. Two of 108 pooled samples showed a faint immuno reaction in the anti-NSE and anti-GFAP immunoblot. Further two particles showed a faint reaction for NSE and GFAP in immuno histochemistry, thus suggesting the presence of CNS. Even though CNS tissue could not be shown in the histological investigation, we used our findings to estimate the worst case scenario for human BSE exposure risk (HER) by lung contaminated by CNS emboli. The content of CNS in the samples was estimated to be about 0.11% when the respective immuno reactions were calibrated against standards containing known brain concentrations. Under the assumption that only one lung in the pooled samples was contaminated with BSE-infected central nervous tissue, the HER was calculated to reach a maximum of 2.2 x 10(-5) CoID50/consumer after consumption of a sausage with a portion of 10% lung. The results of our study suggest that the contamination of the lung with CNS after using a conventional cartridge-fired stunner cannot be excluded, however, the incidence appears to be very low. In addition, presumed CNS emboli, if at all, are microscopically small. Furthermore the incidence of BSE in Germany is very low and lungs of cattle are usually not consumed. Thus we can judge the potential for human oral exposure after consumption of lungs of cattle which were stunned in Germany to be extremely low. A final assessment, however, is impossible as there is no knowledge about the minimum infectious dose for humans.

Abattoirs↗

PubMed2002

[A clinicopathological and prognostic study of 22 cases central neurocytoma].

OBJECTIVE: To investigate the clinicopathological features and prognosis of 22 cases of central neurocytoma (CNC), representing 0.48% of a series of 4 528 patients undergoing biopsy for central nervous system tumors. METHODS: The histopathological, ultrastructral, immunohistochemical and clinical features of CNC were studied by electron microscopic examination and immunohistochemical stain for Synaptophysin (Syn), neuron special enolase (NSE), Leu-7, glial fibrillary acid protein (GFAP), MBP and proliferating cell nuclear antigen (PCNA). RESULTS: The age of the cases ranged from 4 to 44 (average 27.9 years) with all tumors localized in the ventricles. In the 18 patients followed up, 14 were alive for 8 months to 14 years and 11 months after the operation, and 4 died. The average survival period was 70.7 months. Histologically, the tumor in all 22 cases had the oligodendroglioma-like pattern with honeycomb appearance and cell-free islands of eosinophilic matrix. Cellular anaplasia, mitosis and necrotic areas were rarely seen in the tumors. Immunohistochemical study demonstrated strong positivity for Syn, NSE and Leu-7, and negative for GFAP and MBP. Ultrastructural features showed presence of round tumor cells with abundant cell processes containing microtubules, neurosecretory granules, clear vesicles and lysosome-like structures. CONCLUSIONS: The differential diagnosis between CNC and oligodendroglioma could not be established by routine light microscopy. The importance of immunohistochemical and electron microscopic studies for making a correct diagnosis is emphasized. The prognosis of patients is usually favorable, even if the tumor was resected subtotally. The relationship between the presence of anaplastic histological features in CNC and patient outcome remains unclear.

Adolescent↗

PubMed1992

The combination of gamma-interferon and tumor necrosis factor causes a rapid and extensive differentiation of human neuroblastoma cells.

Neuroblastoma (NB), a tumor originating from the sympathetic nervous system, is the most common extracranial neurological tumor of childhood. Human NB cells may differentiate in vitro under treatment with biological agents, as gamma-interferon (IFN-gamma) and tumor necrosis factor (TNF). Unfortunately, NB cell lines resistant to the differentiation-inducing effects of both drugs have been observed. Here we demonstrate that a combination of IFN-gamma plus TNF causes extensive and generalized differentiation of NB cells toward a neuronal phenotype. Both IFN-gamma and TNF, given alone, moderately reduced cell growth and induced partial morphological maturation. Their combination further reduced cell proliferation. The combined treatment gave a synergistic rather than additive cytostatic effect, documented also by a dramatically enhanced differentiation toward a neuronal morphology. Membrane immunofluorescence showed a homologous and heterologous up-regulation of IFN-gamma receptor, as well as a marked induction of HLA Class I antigens and, to a lesser extent, of Class II antigens on NB cells induced to differentiate. Treatment of NB cell lines with IFN-gamma/TNF results in the induction of a differentiated phenotype, as indicated by the increased expression of the Mr 160,000 and 200,000 neurofilament proteins and that of microtubule-associated proteins. Evaluation of biochemical markers of neuronal differentiation confirmed the ability of the combined treatment to induce neuroblast maturation. These results suggest that the combination of IFN-gamma and TNF should be considered for experimental clinical trials in neuroblastoma.

Acetylcholinesterase↗

PubMed2004

[Studies on the injury effects of hippocampus induced by high power microwave radiation in rat].

OBJECTIVE: To study the changes of morphology and function in rat hippocampus induced by high power microwave (HPM) radiation. METHODS: Fifty male Wistar rats were radiated by HPM. Then their learning and memory abilities were tested with Y maze and were sacrificed 6 h, 1 d, 3 d and 7 d after radiation. The hippocampus was taken out to study the basic pathologic changes, apoptosis and the expressions of neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) by means of HE staining, Nissel body staining, in situ terminal end labeling and immunohistochemistry. RESULTS: The learning and memory abilities of rats reduced significantly after HPM radiation. HPM also resulted in rarefaction, edema and hemangiectasia of hippocampus, nervous cells degeneration and necrosis, decrease or disappearance of Nissel bodies. The injuries were more serious in field CA4 and dentate gyrus, which showed dose-effect relationship, and were progressively aggravated within 7 days. The apoptosis cells were significantly increased. NSE was increased in neurons. The NSE positive areas were also seen in the interstitial matrix and blood vessels. GFAP was increased in astrocytes, which became shorter and thicker. CONCLUSION: HPM can damage the abilities of learning and memory and results in morphologic changes in hippocampus. The major pathologic changes are degeneration, apoptosis and necrosis of neurons and edema in interstitium. NSE and GFAP play an important role in the pathologic process.

Animals↗

PubMed2004

[Markers of inflammation, autoantibodies to neurospecific antigens and outcome in patients with acute ischemic stroke].

To identify biochemical markers for carotid stroke outcome, blood serum levels of inflammation markers (C-reactive protein, orosomucoid, soluble p-selectin) and autoantibodies (AAB) to neurospecific antigens (glial fibrillary acidic protein, neuron specific enolase, S-100 protein) were studied in 27 patients (mean age 64 +/- 6 years) with acute ischemic stroke in inner carotid artery system on day 1-2 and 21 of the disease onset. To day 21, patients with good rehabilitation of neurological functions (group 1) demonstrated a decrease of C-reactive protein and soluble p-selectin concentrations, and unfavorable disease course was associated with a significant (p<0.05) increase of concentrations of these indices. On day 1 and 7, a level of AAT to glial fibrillary acidic protein was higher (p<0.05) in group 1 than in that with minimal rehabilitation and to day 21 it decreased relatively the baseline level. At the same time, patients with minimal rehabilitation had a stable AAT level. On day 7, the AAT level correlated with expression of neurological deficit on day 21 (r=0.510; p=0.019). No stroke-course-dependent differences were found in dynamics of orosomucoid as well as of AAT to neuron specific enolase and S-100 protein levels.

Acute Disease↗

PubMed1986

[Immunohistochemical study of subependymal giant cell tumor associated with tuberous sclerosis].

Subependymal giant cell tumors from four patients associated with tuberous sclerosis were examined by peroxidase antiperoxidase (PAP) immunohistochemistry for neuron specific enolase (NSE), glial fibrillary acidic protein (GFAP) and S-100 protein. The tumor cells were grouped into three morphological types: ganglion-like pyramidal cells, swollen gemistocytic cells and fibrillated spindle cells. There was little difference in immunohistochemical property between these cell types in each case, suggesting that the tumor cells of different cell types have a same origin. The majority of the tumor cells was intensely stained for NSE in every case. On the other hand, for GFAP none of the tumor cells was stained in two cases, and the minority was weakly stained in the other two cases. The intensity of staining of the tumor cells for S-100 protein varied from case to case. Our findings together with those of others show the definite immunohistochemical characteristics of subependymal giant cell tumors with tuberous sclerosis, i.e., strong positivity for NSE in contrast to their negativity or weak positivity for GFAP. However, these findings do not necessarily suggest that the tumor is of neuronal origin, because in the case of neoplasms the significance of the presence of NSE as indicative of neuronal origin is open to question.

Adolescent↗

PubMed1987

Postnatal evolution of the human pineal gland. An immunohistochemical study.

Pineal glands from 16 infants ranging from 38 weeks gestation to 3 years of age were fixed in buffered formalin; Paraffin sections were stained for neuron-specific enolase, glial fibrillary acidic protein, and S-100 protein (S-100) using the peroxidase-antiperoxidase method and hematoxylin and eosin, Masson-Fontana, and Bodian stains. The pineal glands of neonates consisted of cords of closely packed, dark, nucleated cells (type I) with intervening loosely arranged, large, clear cells (type II). The type I cells were frequently pigmented and occasionally exhibited rosette formation. They were positive for S-100 and negative for neuron-specific enolase. The type II cells were strongly positive for neuron-specific enolase and negative for melanin and S-100. The type I cells were the predominant cell type at birth; however, the number of type II cells gradually increased with age, and by the age of 1 year, only scattered S-100 positive cells, consistent with sustentacular cells, were found. The findings indicated that the human pineal glands undergo a remarkable morphologic and functional evolution as an endocrine organ in the postnatal life.

Cell Differentiation↗

PubMed1985

[Analysis of various factors related to the prognosis of medulloblastoma].

Thirty-three patients with medulloblastoma were treated in our department by surgical resection of tumor and radio-chemotherapy, and obtained 40.1% of 5 year survival. The present study analyzed the various factors related to the prognosis from clinical and pathological points of view in our series. Good factors were extensive resection of tumor and completement of whole CNS radiationtherapy. In contrast, poor factors were the presence of subarachnoid dissemination of tumor and identification of differentiated cells in tumor tissue histologically. The protection and treatment of subarachnoid dissemination was difficult with any kind of therapy. With surgical specimens obtained from 26 patients, glial fibrillary acidic protein (GFAP) and neuron specific enolase (NSE) were stained immunohistochemically. Two cases out of 12 with GFAP and/or NSE positive cells are alive and their mean survival time is 27 months. On the other hand, eight cases out of 14 without GFAP nor NSE positive cells are alive and their mean survival time is 44 months.

Adolescent↗

PubMed1993

Immunohistochemical study of intracranial cysts.

We present the immunohistochemical study of 11 cases of intracranial cysts: two extraventricular ependymal cysts, three colloid cysts of the third ventricle, four extraventricular choroidal cysts and two Rathke's cleft cysts. Antibodies against glial fibrillary acidic protein (GFAP), cytokeratins (AE1, CK5D, AE3), S-100 protein, epithelial membrane antigen (EMA), vimentin, neuron specific enolase (NSE), neurofilaments protein (NF) and prealbumin, were used. The epithelium of choroidal cysts, showed strong immunoreactivity for Prealbumin and cytokeratins, similar to the normal choroid plexus epithelium. The ependymal cysts showed epithelial immunoreactivity for GFAP and S-100, both glial markers expressed by the normal ependymal epithelium. On the contrary, the epithelial wall of colloid cysts and Rathke's cleft cyst, expressed epithelial markers (cytokeratins and EMA) but no neuroepithelial markers, with a immuno-phenotype similar to that of other cysts of endodermal nature. This finding supports the neuroepithelial origin for choroid and ependymal cysts, and an endodermal nature for colloid and Rathke's cleft cysts. We conclude that these immunohistochemical markers are useful in the differential diagnosis of intracranial cysts.

Brain Diseases↗

PubMed1995

Immunohistochemical and ultrastructural studies of stromal cells in hemangioblastoma.

In order to shed more light on the controversial tissue histogenesis of the stromal cells (SC), light microscopic, immunohistochemical and electron microscopic studies were performed on surgical specimens of hemangioblastomas (36, 26 and 7 cases, respectively). SC were immunoreactive for vimentin, S-100 protein, and neuron specific enolase (NSE) in all cases. Occasional SC were also positive for desmin, smooth muscle actin, Factor VIII, Ulex europaeus lectin receptors, GFAP, and Factor XIIIa. However, majority of these cells were negative with all the endothelial and smooth muscle cell markers used. Electron microscopy demonstrated several different types of SC that were reminiscent of pericytes, smooth muscle cells and abnormal endothelium as well as the intermediate forms between all the above cell types. Few SC were found lining the vascular lumina. Some SC formed small cavities reminiscent of early capillaries. However, typical Weibel-Palade bodies were not found in these SC. It is concluded that SC represents a heterogeneous population of lipidized cells, derived predominantly from the vasogenic mesenchyme. Although immunohistochemistry failed to reveal any consistent antigenic property of SC, ultrastructural findings strongly support the hypothesis that these cells are modified or abnormally differentiated endothelial cells and pericytes.

Adolescent↗

PubMed1998

[Central neurocytoma].

INTRODUCTION: A central neurocytoma (CN) is a rare tumor, of neuronal origin, well-differentiated and found intraventricularly. It mainly affects young adults. Firm diagnosis is made on immunohistochemical (IHQ) and ultrastructural studies, since on optic microscopy it is similar in appearance to an oligodendroglioma or to an ependymoma. PATIENTS AND METHODS: We studied 4 cases, three after surgical resection and one on autopsy. The average age was 29, ranging from 3 to 63. Both sexes were equally affected. In all cases IHQ techniques were used (GFAP, neurofilament, synaptophysin and specific neuronal enolase) and they were studied by electron microscopy. RESULTS: IHQ was negative for GFAP and neurofilament, but intensely positive for synaptophysin and specific neuronal enolase. On ultrastructural study there were few neurofilaments, microtubules and dense central granules typical of neural differentiation. CONCLUSIONS: The findings in our cases lead to diagnosis of NC and confirm that this tumor is a distinct clinicopathological entity.

Adolescent↗

PubMed1995

Non-neuronal enolase is an endothelial hypoxic stress protein.

The hypoxia-associated proteins (HAPs) are five cell-associated stress proteins (M(r) 34, 36, 39, 47, and 57) up-regulated in cultured vascular endothelial cells (EC) exposed to hypoxia. While hypoxic exposure of other cell types induces heat shock and glucose-regulated proteins, EC preferentially up-regulate HAPs. In order to identify the 47-kDa HAP, protein from hypoxic bovine EC lysates was isolated, digested with trypsin, and sequenced. Significant identity was found with enolase, a glycolytic enzyme. Western analyses confirmed that non-neuronal enolase (NNE) is up-regulated in hypoxic EC. Western analysis of subcellular fractions localized NNE primarily to the cytoplasm and confirmed that it was up-regulated 2.3-fold by hypoxia. Interestingly, NNE also appeared in the nuclear fraction of EC but was unchanged by hypoxia. Northern analyses revealed that NNE mRNA hypoxic up-regulation began at 1-2 h, peaked at 18 h, persisted for 48 h, and returned to base line after return to 21% O2 for 24 h. Hypoxia maximally up-regulated NNE mRNA levels 3.4-fold. While hypoxic up-regulation of NNE may have a protective effect by augmenting anaerobic metabolism, we speculate that enolase may contribute to EC hypoxia tolerance through one or more of its nonglycolytic functions.

Amino Acid Sequence↗

PubMed1993

Human lung enolase: cloning and sequencing of cDNA and its inducibility with dexamethasone.

Corticosteroids used orally and intravenously lead to lung diseases and vascular disorder. To investigate whether enolase levels are also changed by treatment with synthetic steroid dexamethasone (as alteration in the enolase levels have been correlated with lung cancer) we performed the following studies. A cDNA library was prepared from poly(A) mRNA extracted from human lung fibroblast cells. cDNA clone HLE1 containing 1.7 kb insert coding for enolase was isolated. Its identity was confirmed by (a) translation of the hybrid selected mRNA and (b) nucleotide sequence analysis of the insert and comparison with known enolase sequences from other species. The lung enolase is coded by a polypeptide of 458 amino acid residues (mr = 49.5 kD). Nucleotide sequencing and derived amino acid sequence data suggest that the cloned enolase is non-neuronal isoform of enolase (NNE). In lung fibroblast cells, dexamethasone caused remarkable increase in the abundance of the enolase mRNA, which was concentration and time dependent. The induction by dexamethasone required de novo RNA synthesis but not de novo protein synthesis.

Amino Acid Sequence↗

PubMed2003

Molecular cloning of an alpha-enolase from the human filarial parasite Onchocerca volvulus that binds human plasminogen.

Enolase represents a multifunctional protein involved in basic energy metabolism and plasminogen binding and activation at the surface of prokaryotic pathogens. A complete cDNA of 1615 bp of an alpha-enolase from Onchocerca volvulus (Ov-ENO) was isolated using a PCR-based approach. The open reading frame encoded for 435 amino acids and the high degree of conservation included the crucial amino acid residues that participate in the formation of the catalytic site, Mg(2+) binding site, and a hydrophobic motif reported to relate to surface expression. A 1089-bp fragment was expressed in a N-terminal 6 x His-tag expression vector in Escherichia coli. By immunohistological analysis using anti-Ov-ENO rabbit antibodies, native enolase could be detected in most tissues of adult O. volvulus, microfilariae, and infective larvae. Intense staining was observed in the muscles, where the energy consumption is high. The purified recombinant protein fragment revealed plasminogen binding activity in a blot-overlay assay employing anti-plasminogen antibodies. In sera from individuals infected with O. volvulus, IgG antibodies reactive with recombinant Ov-ENO were demonstrated by immunoblot and enzyme-linked immunosorbent analyses. The plasminogen-binding property of O. volvulus alpha-enolase may support plasmin-mediated proteolysis including degradation of host's extracellular matrix thereby promoting the migration of larval stages through tissues. The recognition by antibodies in sera of O. volvulus-infected persons indicate an involvement of the protein in the interaction between the parasite and the human host.

Amino Acid Sequence↗

PubMed2005

Novel role for gabapentin in neuroprotection of central nervous system in streptozotocine-induced diabetic rats.

AIM: To investigate the effect of gabapentin on neural [neuron-specific enolase (NSE)] and glial markers [glial fibrillary acidic protein (GFAP) and S100B] in different brain regions of diabetic rats. METHODS: Diabetes was induced by a single intraperitoneal injection of streptozotocine (50 mg/kg body weight). Rats in one diabetic group received gabapentin (50 mg.kg(-1).d(-1)) and rats in the other diabetic group received vehicle only for 6 weeks. The levels of GFAP, S100B, and NSE were determined by immunoblotting in the hippocampus, cortex, and cerebellum. Lipid peroxidation (LPO as malondialdehyde+ 4-hydroxyalkenals) and glutathione (GSH) levels were also determined in the same brain parts. RESULTS: Total and degraded GFAP content and S100B protein expression in different areas of brain tissues significantly increased in diabetic rats compared to control rats. Similarly, NSE levels were also significantly elevated in hyperglycemic rats. In addition, there was a significant increase in LPO levels in the diabetic rat brain compared to control rat brains. Pretreatment with gabapentin prevented the upregulation of GFAP, S100B, and NSE in all brain regions of diabetic rats. The level of LPO was reduced, but not completely halted, by treatment with gabapentin. CONCLUSION: These results suggest that diabetes causes glial and neuronal injury, possibly as a result of elevated oxidative stress, and that gabapentin protects neurons and glial cells. Thus, we predict that gabapentin treatment will attenuate the hippo-campal and cortical neurodegeneration observed during diabetes mellitus in rats.

Amines↗

PubMed1998

cDNA cloning and characterization of neuron-specific enolase from chicken.

Chicken gamma-enolase cDNA was cloned and its sequence and tissue-specific expression were analyzed. The cDNA, consisting of 2273 bp of nucleotides, was composed of 86 bp of the 5'-noncoding region, 1305 bp of an open reading frame encoding a protein of 434 amino acids and 882 bp of the 3'-noncoding region. The deduced amino acid sequence showed higher homology (more than 90%) to those of mammalian gamma-enolases than to those of chicken alpha- or beta-enolases. Northern blot analysis has revealed that the mRNA for gamma-enolase (2.3 kb) is expressed in the brain and, to much less but significant extents, in the pituitary and adrenal gland of the chicken.

Amino Acid Sequence↗

PubMed2006

Evolution of enzymatic activities in the enolase superfamily: L-fuconate dehydratase from Xanthomonas campestris.

Many members of the mechanistically diverse enolase superfamily have unknown functions. In this report we use both genome (operon) context and screening of a library of acid sugars to assign the L-fuconate dehydratase (FucD) function to a member of the mandelate racemase (MR) subgroup of the superfamily encoded by the Xanthomonas campestris pv. campestris str. ATCC 33913 genome (GI:21233491). Orthologues of FucD are found in both bacteria and eukaryotes, the latter including the rTS beta protein in Homo sapiens that has been implicated in regulating thymidylate synthase activity. As suggested by sequence alignments and confirmed by high-resolution structures in the presence of active site ligands, FucD and MR share the same active site motif of functional groups: three carboxylate ligands for the essential Mg2+ located at the ends of the third, fourth, and fifth beta-strands in the (beta/alpha)7beta-barrel domain (Asp 248, Glu 274, and Glu 301, respectively), a Lys-x-Lys motif at the end of the second beta-strand (Lys 218 and Lys 220), a His-Asp dyad at the end of the seventh and beta-strands (His 351 and Asp 324, respectively), and a Glu at the end of the eighth beta-strand (Glu 382). The mechanism of the FucD reaction involves initial abstraction of the 2-proton by Lys 220, acid catalysis of the vinylogous beta-elimination of the 3-OH group by His 351, and stereospecific ketonization of the resulting enol, likely by the conjugate acid of Lys 220, to yield the 2-keto-3-deoxy-L-fuconate product. Screening of the library of acid sugars revealed substrate and functional promiscuity: In addition to L-fuconate, FucD also catalyzes the dehydration of L-galactonate, D-arabinonate, D-altronate, L-talonate, and D-ribonate. The dehydrations of L-fuconate, L-galactonate, and D-arabinonate are initiated by abstraction of the 2-protons by Lys 220. The dehydrations of L-talonate and D-ribonate are initiated by abstraction of the 2-protons by His 351; however, protonation of the enediolate intermediates by the conjugate acid of Lys 220 yields L-galactonate and D-arabinonate in competition with dehydration. The functional promiscuity discovered for FucD highlights possible structural mechanisms for evolution of function in the enolase superfamily.

Amino Acid Substitution↗