bromochloroacetic-acid and Astrocytoma

Clear cell meningioma (CCM) is a rare variant of meningioma, which is important to distinguish because of its aggressive behaviour. Sixty-eight cases have been previously described in the literature. In this retrospective study, we report seven cases of CCM operated in our institution between 1994 and 2008.. Seven CCM cases were retrieved from the files of our pathology department. Clinical and radiological data were reviewed. A standard histological study was realized and immunohistochemistry was performed with epithelial membrane antigen (EMA), cytokeratin KL1, progesterone receptors, Ki-67 (MIB-1), S100 protein.. Patients' age ranged from 2 to 70 years (median age: 36 years), with a female predominance (5/7 patients). Three patients belonged to the same family, probably affected by neurofibromatosis type 2. CCM occurred in various locations: medullary region (two), sphenoid wing (two), ponto-cerebellar angle (two), tentorium (one). The tumour could be fully resected in three cases. Follow-up ranged from 3 months to 15 years: recurrence occurred in four patients, three of whom eventually died from the disease.. In our series, the frequency of CCM (0,6% of all meningiomas operated on in our institution) and its histological aspects are almost identical to those observed of the literature. We discuss the predictive value of proliferation index (MIB-1) and the role of patient status and quality of surgical resection in the evolution.. Our study supports the fact that MCC course is less favourable than meningioma WHO grade I, even in the absence of anaplastic area, high mitotic activity, or necrosis. In this series, MIB-1 index was of no interest identifying patients with or without recurrence.

Topics: Adult; Aged; Astrocytoma; Biomarkers, Tumor; Child, Preschool; Diagnostic Errors; Ependymoma; Female; Humans; Keratins; Ki-67 Antigen; Male; Meningeal Neoplasms; Meningioma; Middle Aged; Mucin-1; Neoplasm Proteins; Neurofibromatosis 2; Receptors, Progesterone; Retrospective Studies; S100 Proteins; Young Adult

In contrast to the relatively common soft tissue form of granular cell tumor (GCT), intercerebral GCTs are rare neoplasms. The Schwann cell is the accepted cell of origin for soft tissue GCTs. However, the origin of intracerebral tumors is controversial. We report a case of a GCT intimately associated with an anaplastic astrocytoma. Immunohistochemical staining with glial fibrillary acidic protein demonstrated focal positive staining within the granular cells. Six GCTs from other body sites were stained with glial fibrillary acidic protein for comparison and all were negative. The granular cell component was diffusely positive for S-100 and negative for epithelial membrane antigen and cytokeratin. Ultrastructurally, filaments characteristic of astrocytic cells were demonstrated within some granular cells. Based on our light microscopic, electron microscopic, and immunohistochemical findings, the granular cell component of this anaplastic astrocytoma is likely astrocytic in origin. We propose that these tumors be designated astrocytic neoplasms with granular cell differentiation and their prognoses dictated by the grade of the glial component.

Topics: Astrocytoma; Glial Fibrillary Acidic Protein; Granular Cell Tumor; Humans; Keratins; Male; Middle Aged; S100 Proteins

Cytoplasmic polyadenylation element binding protein 4 (CPEB4) is a regulator of gene transcription and has been reported to be associated with biological malignancy in cancers. However, it is unclear whether CPEB4 has any clinical significance in patients with astrocytic tumors, and mechanisms that CPEB4 contribute to progression of astrocytic tumors remain largely unknown. Here, correlation between CPEB4 expression and prognosis of patients with astrocytic tumors were explored by using qPCR, WB and IHC, and X-tile, SPSS software. Cell lines U251 MG and A172 were used to study CPEB4's function and mechanisms. Co-immunoprecipitation, mass spectrometry, immunofluorescent assay, and western blot were performed to observe the interaction between CPEB4 and Vimentin. CPEB4 mRNA and protein levels were markedly elevated in 12/12 astrocytic tumors in comparison to paratumor. High expression of CPEB4 was significantly correlated with clinical progressive futures and work as an independent adverse prognostic factor for overall survival of patients with astrocytic tumors (relative risk 4.5, 95 % CI 2.1-11.2, p = 0.001). Moreover, knockdown of CPEB4 in astrocytic tumor cells inhibited their proliferation ability , clonogenicity, and invasiveness. Five candidate proteins, GRP78, Mortalin, Keratin, Vimentin, and β-actin, were identified, and the interaction between CPEB4 and Vimentin was finally confirmed. Downregulation of CPEB4 could reduce the protein expression of Vimentin. Our studies first validated that CPEB4 interacts with Vimentin and indicated that high CPEB4 expression in astrocytic tumors correlates closely with a clinically aggressive future, and that CPEB4 might represent a valuable prognostic marker for patients with astrocytic tumors.

Topics: Actins; Adult; Aged; Astrocytoma; Biomarkers, Tumor; Cell Proliferation; Endoplasmic Reticulum Chaperone BiP; Female; Gene Expression Regulation, Neoplastic; Heat-Shock Proteins; HSP70 Heat-Shock Proteins; Humans; Keratins; Male; Middle Aged; Mitochondrial Proteins; Neoplasm Invasiveness; Prognosis; RNA-Binding Proteins; RNA, Messenger; Survival Analysis; Vimentin

Metastatic carcinoma, which is a common malignant tumor seen in the central nervous system is often difficult to distinguish from glioblastoma multiforme. In general, neoplastic cells maintain fidelity in the expression of parent cell intermediate filament and immunohistochemistry remains the mainstay in diagnosis. A panel consisting of GFAP (usually positive for astrocytic tumors) and cytokeratin (usually positive for metastatic carcinomas) is most commonly used for this purpose. However, co-expression of two or more classes of intermediate filament proteins by neoplasms is a widespread phenomenon and there are reports of glial neoplasms expressing keratin markers. Our aims and objectives were to analyse the expression of both cytokeratin and GFAP in different glial tumors and metastatic carcinomas. Cases were collected for a period of two years. All the cases were diagnosed as primary or metastatic intracranial tumors. Formalin-fixed paraffin-embedded thin sections were taken on egg-albumin coated slides and immunostaining with GFAP and polyclonal cytokeratin was done. Forty-five tumors were analysed, including 35 glial neoplasms and 10 metastatic carcinomas of which 7 of the 32 astrocytic neoplasms (22%) showed focal immunoreactivity with pancytokeratin. All of the glial tumors but none of the metastatic carcinomas were positive with GFAP. So our conclusion was that co-expression of GFAP and CK is a fairly common phenomenon, especially in case of undifferentiated and high grade gliomas and this must be kept in mind while differentiating these cases from metastatic carcinoma, as CK positivity does not rule out the diagnosis of a glial neoplasm. Further studies with an expanded panel of CK is most useful for this.

Topics: Astrocytoma; Biomarkers, Tumor; Brain Neoplasms; Carcinoma; Diagnosis, Differential; Glial Fibrillary Acidic Protein; Glioblastoma; Glioma; Humans; Immunohistochemistry; Keratins; Oligodendroglioma

Exposure of cells to stress, particularly oxidative stress, leads to misfolding of proteins and, if they are not refolded or degraded, to cytoplasmic protein aggregates. Protein aggregates are characteristic features of a variety of chronic toxic and degenerative diseases, such as Mallory bodies (MBs) in hepatocytes in alcoholic and non-alcoholic steatohepatitis, neurofibrillary tangles in neurons in Alzheimer's, and Lewy bodies in Parkinson's disease. Using 2D gel electrophoresis and mass spectrometry, we identified p62 as a novel MB component. p62 and cytokeratins (CKs) are major MB constituents; HSP 70, HSP 25, and ubiquitinated CKs are also present. These proteins characterize MBs as a prototype of disease-associated cytoplasmic inclusions generated by stress-induced protein misfolding. As revealed by transfection of tissue culture cells overexpressed p62 did not induce aggregation of regular CK filaments but selectively bound to misfolded and ubiquitinated CKs. The general role of p62 in the cellular response to misfolded proteins was substantiated by detection of p62 in other cytoplasmic inclusions, such as neurofibrillary tangles, Lewy bodies, Rosenthal fibers, intracytoplasmic hyaline bodies in hepatocellular carcinoma, and alpha1-antitrypsin aggregates. The presence of p62 along with other stress proteins and ubiquitin in cytoplasmic inclusions indicates deposition as aggregates as a third line of defense against misfolded proteins in addition to refolding and degradation.

Topics: Adaptor Proteins, Signal Transducing; alpha 1-Antitrypsin Deficiency; Animals; Astrocytoma; Central Nervous System Neoplasms; CHO Cells; Cricetinae; Heat-Shock Proteins; Humans; Inclusion Bodies; Keratins; Liver Diseases; Mice; Neurodegenerative Diseases; Protein Binding; Protein Folding; Proteins; Sequestosome-1 Protein; Stress, Physiological; Ubiquitin

A diagnostic problem can occur at the time of intraoperative consultation of neurosurgical tumors as to whether the tumor is of neuroectodermal origin or whether it represents an epithelial metastasis from another site. Intraoperative diagnoses based on hematoxylin and eosin stained frozen sections are often later confirmed by immunocytochemical analysis of formalin-fixed, paraffin-embedded tissue sections that are not available at the time of surgery. The objective of the current study was to demonstrate that the application of direct immunofluorescence to the intraoperative diagnosis of neurosurgical tumors would provide unequivocal, and nearly immediate results. This report describes a new application of an existing technique for an optimized, rapid procedure utilizing direct immunocytochemistry with fluorescence-labeled primary antibodies to analyze surgical biopsies intraoperatively. The examination of five neurosurgical biopsies established a neuroectodermal origin of three tumors via immunolabeling for glial fibrillary acidic protein (GFAP) and lack of labeling with keratin markers, whereas several metastatic lung carcinomas were identified by immunostaining for keratin, but not GFAP, markers. The results of the direct immunolabeling method were unequivocal and required only minutes. The same diagnoses were confirmed by standard immunocytochemical labeling of formalin-fixed, paraffin-embedded sections, though it required several days to obtain the results. Direct immunofluorescence using fluorescently conjugated primary antibodies is a practical and rapid method for deciding whether a neurosurgical tumor is a primary glial or an epithelial metastatic tumor in origin. It is the first reported application of the technique for this aspect of rapid neurosurgical diagnosis.

Topics: Antibodies, Monoclonal; Astrocytoma; Brain Neoplasms; Fluorescent Antibody Technique, Direct; Glial Fibrillary Acidic Protein; Humans; Immunoenzyme Techniques; Intraoperative Period; Keratins; Neurosurgery; Paraffin Embedding

Intracranial nervous-system tumours were diagnosed in three of 1092 bovine necropsy specimens submitted to the Department of Veterinary Pathology, Obihiro University between April 1983 and March 1996. A fourth case was a referral from the Department of Veterinary Pathology, Rakuno Gakuen University. Histopathological examination revealed four types of tumour: intracranial malignant peripheral nerve sheath tumour (MPNST), choroid plexus papilloma, differentiated fibrillary astrocytoma and anaplastic (malignant) astrocytoma. Immunohistochemically, the intracranial MPNST was strongly positive for S-100 protein and vimentin, and in places weakly positive for glial fibrillary acid protein (GFAP). The choroid plexus papilloma was strongly positive for epithelial membrane antigen (EMA), keratin, S-100 protein and vimentin, and positive for GFAP in places. The cytoplasm and fibrous component in the differentiated fibrillary astrocytoma were strongly positive for S-100 protein and GFAP. The anaplastic (malignant) astrocytoma was strongly positive for vimentin, S-100 protein and keratin in the cytoplasm and fibrous processes, and weakly positive for GFAP and EMA in places. Myelin basic protein (MBP) and synaptophysin showed a weak positive reaction in the marginal areas of the tumour.

Topics: Animals; Astrocytoma; Brain Neoplasms; Cattle; Cattle Diseases; Glial Fibrillary Acidic Protein; Glioblastoma; Glioma; Immunohistochemistry; Keratins; Mucin-1; Myelin Basic Protein; Nerve Sheath Neoplasms; S100 Proteins; Vimentin

Several studies have shown that immunoenzymatic staining of formalin-fixed, paraffin-embedded astrocytomas with keratin antibodies frequently yields positive labelling, but no biochemical evidence of keratin expression in astrocytomas has been reported. We have investigated the presence of keratin in astrocytoma and normal brain tissues both by immunofluorescence on frozen sections and by 1D and 2D immunoblotting using seven monoclonal antibodies that, collectively, recognize most keratin polypeptides. Four of these antibodies did not stain neural tissues by immunofluorescence and were also negative by immunoblotting. The remaining three keratin antibodies stained normal brain and/or a high proportion of astrocytomas. Two of these three antibodies only stained glial fibrillary acidic protein (GFAP)-positive cells, while the third only stained GFAP-negative cells. 1D and 2D immunoblotting analysis showed that positive immunofluorescence staining of normal brain and/or astrocytomas seen with these three keratin antibodies was due to cross-reactivity with non-keratin proteins, such as GFAP. These results demonstrate that, contrary to earlier suggestions, keratin polypeptides are not frequently expressed in astrocytomas. Our studies also emphasize that keratin antibodies should be used cautiously for the differential diagnosis of undifferentiated gliomas from tumours of non-glial origin.

Topics: Antibodies, Monoclonal; Astrocytoma; Brain; Brain Neoplasms; Cross Reactions; False Positive Reactions; Fluorescent Antibody Technique, Direct; Glial Fibrillary Acidic Protein; Humans; Immunoblotting; Keratins

We examined the expression of glial- and neuronal-specific mRNAs within human gliomas using in situ hybridization. We found that low-grade astrocytomas contained a high number of proteolipid protein (PLP) mRNA-positive cells and that the number of PLP-stained cells decreased markedly with increasing tumor grade. Interestingly, the ratio of PLP mRNA-stained cells:myelin basic protein (MBP) mRNA-stained cells in normal white matter and low-grade astrocytoma was about 2:1 but approached 1:1 with increasing tumor grade. This parameter appeared to be a good indicator of tumor infiltration in astrocytomas, so we tested this in the analysis of other gliomas. Unlike astrocytomas, oligodendrogliomas were found consistently to contain few PLP mRNA- or MBP mRNA-expressing cells. In contrast, gemistocytic astrocytomas, typically highly invasive tumors, contained high numbers of PLP-positive cells and a ratio of PLP mRNA:MBP mRNA-stained cells of about 1.5:1, similar to low-grade astrocytomas. Nonradioactive in situ hybridization also enabled the morphological identification of specific cells. For example, gemistocytic astrocytes, which were found to be strongly vimentin mRNA positive, contained little glial fibrillary acidic protein mRNA and did not stain for PLP or MBP mRNAs. Neuronal mRNAs, such as neurofilament 68, were observed in small numbers of entrapped neurons within gliomas but were uninformative with respect to predicting tumor grade. Our results suggest that oligodendrocytes survive low-grade tumor infiltration and that glial tumor cells, unlike cell lines derived from them, do not express oligodendrocyte or neuronal mRNAs. In addition, the expression of mRNAs for the two major myelin protein genes, PLP and MBP, could be used to predict the grade and extent of tumor infiltration in astrocytomas.

Topics: Astrocytoma; Brain Neoplasms; Gene Expression Regulation, Neoplastic; Glial Fibrillary Acidic Protein; Glioma; Humans; In Situ Hybridization; Keratins; Myelin Basic Protein; Myelin Proteolipid Protein; Neurofilament Proteins; Neuroglia; Neurons; Oligodendroglia; Oligodendroglioma; RNA, Messenger; RNA, Neoplasm; Vimentin

Topics: Animals; Astrocytoma; Blindness; Brain; Brain Neoplasms; Glial Fibrillary Acidic Protein; Goat Diseases; Goats; Immunohistochemistry; Keratins; Motor Activity; Nerve Tissue Proteins; Neurofilament Proteins; Vimentin

In this study, 29 formalin-fixed, paraffin-embedded astrocytic tumors were analyzed immunocytochemically with the antikeratin monoclonal antibodies Mak-6 and Cam 5.2 and a polyclonal antibody against glial fibrillary acidic protein (GFAP). Immunoreactivity for Mak-6 was present in 29 cases (100%) including six well-differentiated astrocytomas, nine anaplastic astrocytomas, and 14 glioblastomas multiforme. Cam 5.2 immunoreactivity was focally present in one case of GBM (4%) but was absent in the remaining 28 cases. All cases were immunoreactive with an antibody against GFAP. Cytokeratin (CK) expression was examined in extracts of four separate well-characterized astrocytoma cell lines by Western blotting with the monoclonal antibodies Mak-6, Cam 5.2, and anti-CK 18 and by Northern analysis using a cDNA probe for the human CK 18 gene. The Western blots revealed the presence of immunoreactive bands corresponding to CK numbers 14/15, 16, and 18 in extracts from all four cell lines and additional bands corresponding to CK 8 in 3/4 lines and CK 19 in 1/4 lines. Northern analysis detected CK 18 mRNA in extracts from 2/4 astrocytic cell lines. These findings demonstrate that CK immunoreactivity is frequent in astrocytic tumors and confirm through the molecular and biochemical analysis of CK 18 gene expression and of keratin intermediate filament proteins that the basis for CK immunoreactivity in astrocytic tumors is bona fide CK expression, not cross-reactivity with other antigens or artifact. The demonstration of CK expression by astrocytic neoplasms has important implications for pathologists involved in the diagnosis of poorly differentiated tumors.

Topics: Antibodies, Monoclonal; Astrocytoma; Blotting, Northern; Blotting, Western; Glial Fibrillary Acidic Protein; Glioblastoma; Humans; Immunohistochemistry; Keratins; Paraffin Embedding; Tumor Cells, Cultured

The Cavitron ultrasonic surgical aspirator (CUSA) is a dissecting system that allows quick and effective removal of CNS tumors without traction or excessive manipulation of normal tissue. In this article, the immunoperoxidase staining patterns of cytology specimens obtained with the CUSA are compared with those from their corresponding resected surgical specimens employing a battery of monoclonal and polyclonal antibodies. Eleven cases of meningioma, three cases of glioblastoma multiforme, one astrocytoma, and two schwannomas were evaluated. In both CUSA cytologic biopsies and surgical biopsies, all the meningiomas showed strong staining for vimentin and epithelial membrane antigen, while two showed focal staining for cytokeratins. The glioblastoma multiforme and astrocytoma cases showed positivity for vimentin, S-100 protein, and glial fibrillary acidic protein, while the schwannomas stained positively for vimentin and S-100 protein. With only rare exceptions, the immunocytochemistry of the CUSA and surgical specimens correlated well in all of these cases in terms of strength of reaction and localization. There were no false-positive staining reactions in the CUSA material. This study suggests that reliable morphologic and immunoperoxidase studies can be performed on cytologic material obtained by the CUSA, which could aid in making an accurate and specific diagnosis of a variety of CNS tumors.

Topics: Astrocytoma; Biopsy; Cytodiagnosis; Glial Fibrillary Acidic Protein; Glioblastoma; Humans; Immunoenzyme Techniques; Keratins; Meningioma; Neoplasms, Nerve Tissue; Neurilemmoma; Neurosurgery; S100 Proteins; Suction; Ultrasonic Therapy; Vimentin

Immunohistochemical analysis of 30 paraffin-embedded astrocytic neoplasms was performed to correlate the expression of intermediate filament proteins with histologic subtype. Each tumor was studied with monoclonal antibodies to keratin, vimentin, desmin, 200-kd neurofilament protein, and glial fibrillary acidic protein (GFAP). Immunoreactivity with the anti-keratin monoclonal antibodies AE1 and AE3 was demonstrated in 24 cases (80%) including 4 of 6 (66%) well-differentiated astrocytomas (WDAs), 10 of 12 (83%) anaplastic astrocytomas (ANAs), and 10 of 12 (83%) glioblastomas multiforme (GBMs). These cases were further studied with the monoclonal antikeratin antibodies 34 beta E12 and 34 beta H11. Of the 24 AE1/AE3-positive cases, 14 (58%) reacted with 34 beta E12. None of the cases was reactive with 34 beta H11. Vimentin expression was demonstrated in 24 cases (80%), including 2 of 6 (33%) WDAs, 11 of 12 (92%) ANAs, and 11 of 12 (92%) GBMs. Coexpression of keratin and vimentin was observed in 20 cases (67%), including 2 of 6 WDAs, 9 of 12 (75%) ANAs, and 9 of 12 (75%) GBMs. Immunoreactivity with GFAP antibody was present in all 30 (100%) cases, but none of the tumors was reactive with antibodies to desmin or 200-kd neurofilament protein. These findings demonstrate that expression of both keratin and vimentin intermediate filaments is common in astrocytic neoplasms regardless of histologic grade.

Topics: Astrocytoma; Biomarkers, Tumor; Glial Fibrillary Acidic Protein; Glioblastoma; Humans; Immunoenzyme Techniques; Intermediate Filament Proteins; Keratins; Staining and Labeling; Vimentin

Specimens of neoplastic tissues from 19 dogs and 4 cats were examined immunohistochemically for intermediate filament expression, using commercially available antibodies. Staining was observed in a wide range of tumor tissues and in normal internal controls by use of antibodies to vimentin, desmin, glial fibrillary acidic protein, and low and high molecular weight cytokeratins. Intermediate filament expression was found to be consistent with light and/or electron microscopic findings, and hence believed to be an accurate indicator of tumor histogenesis in cats and dogs. Three fixatives were evaluated for their relative abilities to preserve antigenicity. Absolute alcohol was superior to B5 fixative and both were superior to formalin. Some tissues that clearly displayed intermediate filament antigens with alcohol and B5 fixative failed to stain when fixed in formalin.

Topics: Adenocarcinoma; Animals; Astrocytoma; Cat Diseases; Cats; Cytoskeleton; Desmin; Dog Diseases; Dogs; Immunohistochemistry; Intermediate Filament Proteins; Intermediate Filaments; Keratins; Leiomyosarcoma; Melanoma; Neoplasms; Neurilemmoma

Monoclonal antibodies (AE1/3, CAM 5.2 and PKK-1) and polyclonal antisera against the cytokeratin proteins were reacted with a range of astrocytic tumours, oligodendrogliomas and ependymomas. Seven of 12 cases (58%) of glioblastoma multiforme, five of eight (63%) anaplastic astrocytomas and two of five (40%) well-differentiated astrocytomas were immunoreactive with AE1/3 but not with the other anti-cytokeratin antibodies. In oligodendrogliomas, AE1/3 stained isolated astrocyte-like cells as well as scattered neoplastic oligodendrocytes in four of eight cases (50%) cases. Four ependymomas were negative for all cytokeratin markers examined. The immunostaining of astrocytomas and oligodendrogliomas with AE1/3 might represent co-expression of cytokeratin with glial fibrillary acidic protein by gliomas and calls for caution in the use of these antibodies in the differential diagnosis between gliomas and carcinomas.

Topics: Adult; Antibodies, Monoclonal; Astrocytoma; Child; Glioblastoma; Humans; Immunohistochemistry; Keratins; Oligodendroglioma

The successful application of immunostaining on smear preparations from 34 tumours of the central nervous system using the peroxidase-antiperoxidase technique and the avidin-biotin-peroxidase complex technique with commercially available antibodies is described. When combined with conventional smear preparations, the technique contributed to a rapid and accurate diagnosis in 79% of cases and is advocated to be a useful adjunct in neurosurgical diagnosis.

Topics: Antigens; Astrocytoma; Biopsy; Brain Neoplasms; Glial Fibrillary Acidic Protein; Humans; Immunohistochemistry; In Vitro Techniques; Keratins; Membrane Glycoproteins; Meningioma; Mucin-1; S100 Proteins; Spinal Cord Neoplasms