Plasma amino acids indicate glioblastoma with ATRX loss.

Glioblastoma (GB) is the most common primary brain tumour in adults. The lack of molecular biomarker, non-specific symptoms and fast growth rate often result in a significant delay in diagnosis. Despite multimodal treatment, the prognosis remains poor. Here, we verified the hypothesis that amino acids (AA) regulating the critical metabolic pathways necessary for maintenance, growth, reproduction, and immunity of an organism, may constitute a favourable target in GB biomarker research. We measured the plasma amino acids levels in 18 GB patients and 15 controls and performed the quantitative and qualitative metabolomic analysis of free AA applying high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS). We present both the raw data and the results of our statistical analysis. The majority of AA were lowered in the study group in comparison to the control group. Five of these (arginine, glutamic acid, glutamine, glycine, and histidine) differed significantly (all p < 10-5 and AUC > 0.9). Plasma levels of leucine and phenylalanine decreased in the case of GB with lost alpha-thalassemia/mental retardation X-linked (ATRX) expression on immunohistochemistry (p = 0.003 and 0.045, respectively). We demonstrated for the first time that certain plasma-free AA levels of GB patients were significantly different from those in healthy volunteers. Target profiling of plasma-free AA, identified utilizing LC-QTOF-MS, may present prognostic value by indicating GB patients with lost ATRX expression. The on-going quest for glioma biomarkers still aims to determine the detailed metabolic profile and evaluate its impact on therapy and prognosis.

Calcifying pseudoneoplasm of the foramen magnum--Case report and review of the literature.

We present a case of a 29-year-old male with a calcifying pseudoneoplasm of the neuraxis (CAPNON) located in the region of the foramen magnum, treated successfully by complete resection. After a 2-year follow-up the patient remains recurrence free. Clinical and histopathological characterization of CAPNON is provided with special emphasis on the intraoperative and neuroradiological features of the lesion.

Isolated neurosarcoidosis mimicking intracranial tumours - Analysis of 3 cases.

Isolated neurosarcoidosis (INS), as a disease of low prevalence, is commonly overlooked in differential diagnosis, and its discovery on histopathological examination usually comes as a surprise. Preoperative diagnosis is difficult because the clinical picture of INS is non-specific. Its symptoms depend on the location of the lesions, and the MRI results are similar to those found in meningiomas or optic nerve gliomas. Although up to 5% of all sarcoidosis patients present with neurological symptoms, those with INS are exceptionally infrequently encountered. Three cases of INS are presented here, analysing their clinical course and radiological images, in order to determine characteristic traits that might lead to a correct diagnosis.

Astrocytoma-associated antigens - IL13Rα2, Fra-1, and EphA2 as potential markers to monitor the status of tumour-derived cell cultures in vitro.

BACKGROUND: The molecular heterogeneity of high-grade astrocytomas underlies the difficulties in the development of representative and valuable in vitro experimental models for their studies. The purpose of our study was to estimate the value of astrocytoma-associated antigens (AAAs) - IL13Rα2, Fra-1, EphA2 - and the most common molecular aberrations typical for astrocytomas as potential markers to screen the status of tumour-derived cell cultures in vitro. METHODS: The tumour-derived cell cultures were established from high-grade astrocytomas. The expression analyses of the tested genes were performed via semi-quantitative real-time PCR and subsequently verified by immunohistochemical and immunocytochemical technique. The analyses of molecular aberrations at DNA level included gene dosage status evaluation based on real-time PCR, sequencing analysis, and loss of heterozygosity (LOH) assay. RESULTS: The expression analyses based on semi-quantitative real-time PCR showed that in the final stage of culture the expression level of all tested AAAs was significantly higher or at least comparable to that of primary tumours; however, two expression patterns were observed during cell culture establishment. Analysis at the single cell level via immunocytochemistry also demonstrated an increase of the level of tested proteins and/or selection of tumour cell populations strongly positive for AAAs vs. other cell types including admixed non-tumoural cells. Confrontation of AAA expression data with the results of molecular analyses at DNA level seems to support the latter, revealing that the expression pattern of astrocytoma-associated antigens in tumour-derived cells in subsequent stages of culture is convergent with changes in the molecular profile of examined cell populations. CONCLUSIONS: The consistency of the obtained results seems to support the use of the selected AAAs, in particular IL13Rα2 and Fra-1, as tools facilitating the establishment of tumour-derived cultures. However, the intratumoural heterogeneity of high-grade astrocytomas may require further detailed characterisation of the molecular profile of a tumour in order to evaluate the value of the experimental model in relation to the individual context of particular studies.

Supratentorial neurenteric cyst--a case report.

Supratentorial neurenteric cyst is a rare congenital lesion. We report here a case of a 33-year-old female who presented with seizures. A multicystic lesion in the left premotor cortex with moderate contrast enhancement was demonstrated with MRI. Microscopically, the lesion showed small cystic structures filled with a proteinaceous fluid. The wall of the cysts was lined with a single layer of ciliated columnar or cuboidal epithelium on a basement membrane. Glandular structures resembling gastrointestinal glands were also present. The cells of the cyst lining and glandular structures revealed strongly positive immunoreactions for epithelial markers (cytokeratin and epithelial membrane antigen).

Primary extranodal marginal zone B-cell lymphoma of the mucosa-associated lymphoid tissue type in the central nervous system (MZL CNS) presented as traumatic subdural hematoma and subarachnoid bleeding - case report.

The study describes a very rare case of primary extranodal marginal zone Bcell lymphoma of the central nervous system (MZL CNS) with an unusual clinical and radiological presentation mimicking subarachnoid bleeding and subdural hematoma (SDH) after head injury. The patient presented symptoms which had commenced 3 weeks earlier: a gradually-progressing headache associated with periodic right-sided cramp of the face muscles and numbness of the right upper limb. During urgent craniotomy for drainage of the presumed SDH, a tumor mass histopathologically and immunohistochemically matching marginal zone B-cell lymphoma was found. Molecular analysis confirmed monoclonal immunoglobulin heavy chain gene (IgH) rearrangement; the patient had previously suspected nodal lymphoma because of cervical lymphadenopathy, but histopathological, immunohistochemical and molecular examination excluded malignant lymphoma. The patient underwent successful radiotherapy, and achieved complete response. At present, no evidence of either systemic disease or lymph node enlargement has been found. The recognition of an indolent type of lymphoma in a rare anatomical localization is very important due to the proper management of the patient.

Molecular alterations in meningiomas: association with clinical data.

The aim of our study was to evaluate the frequency of deletions on chromosomes 1, 9, 10, 14, 18 and 22 in 75 benign and 15 atypical meningiomas and correlate them with clinical findings. Paired normal and tumor DNA samples were analyzed for loss of heterozygosity (LOH), using 24 microsatellite markers and PCR techniques. Statistical analysis showed that deletions on chromosomes 14 and 18 were significantly associated with tumor grade of meningiomas (p = 0.048 and p = 0.03, respectively). In addition, we found a marginally increased frequency of LOH on chromosome 9 in atypical meningiomas (p = 0.06). Interestingly, LOH on chromosome 14 was significantly associated with tumor size (p = 0.049), as the risk of developing a tumor of more than 4 cm in diameter was 6-times the risk of developing tumor with diameter below 4 cm. The most frequent genetic abnormality in meningiomas is 22 LOH, which seems to be confirmed by the present study in which high frequency of such abnormality was observed (67%). We found associations between chromosome 22 status and histological subtype. LOH on chromosome 22 was more frequent in fibrous meningiomas than in the meningothelial variant (p = 0.001). Besides that, we found a relationship between 22 LOH status and tumor localization: the frequency of LOH in skull base-localized tumors was significantly lower compared to parasagittal meningiomas (p = 0.0004). Our results indicated that allelic loss on chromosomes 9, 10, 14, 18 and 22 may be associated with meningioma pathogenesis and progression.

Selection of reference genes for gene expression studies in astrocytomas.

This study was aimed to test a panel of six housekeeping genes (GAPDH, HPRT1, POLR2A, RPLP0, ACTB, and H3F) so as to identify and validate the most suitable reference genes for expression studies in astrocytomas. GAPDH was the most stable and HPRT1 was the least stable reference gene. The effect of reference gene selection on quantitative real-time polymerase chain reaction data interpretation was demonstrated, normalizing the expression data of a selected gene of interest. Thus, GAPDH may be recommended for data normalization in gene expression studies in astrocytomas. Nevertheless, a preliminary validation of reference gene stability is required prior to every study.

Glioblastoma-derived spheroid cultures as an experimental model for analysis of EGFR anomalies.

Glioblastoma cell cultures in vitro are frequently used for investigations on the biology of tumors or new therapeutic approaches. Recent reports have emphasized the importance of cell culture type for maintenance of tumor original features. Nevertheless, the ability of GBM cells to preserve EGFR overdosage in vitro remains controversial. Our experimental approach was based on quantitative analysis of EGFR gene dosage in vitro both at DNA and mRNA level. Real-time PCR data were verified with a FISH method allowing for a distinction between EGFR amplification and polysomy 7. We demonstrated that EGFR amplification accompanied by EGFRwt overexpression was maintained in spheroids, but these phenomena were gradually lost in adherent culture. We noticed a rapid decrease of EGFR overdosage already at the initial stage of cell culture establishment. In contrast to EGFR amplification, the maintenance of polysomy 7 resulted in EGFR locus gain and stabilization even in long-term adherent culture in serum presence. Surprisingly, the EGFRwt expression pattern did not reflect the latter phenomenon and we observed no overexpression of the tested gene. Moreover, quantitative analysis demonstrated that expression of the truncated variant of receptor-EGFRvIII was preserved in GBM-derived spheroids at a level comparable to the initial tumor tissue. Our findings are especially important in the light of research using glioblastoma culture as the experimental model for testing novel EGFR-targeted therapeutics in vitro, with special emphasis on the most common mutated form of receptor-EGFRvIII.

Low grade malignant peripheral nerve sheath tumor with mesenchymal differentiation: a case report.

Malignant peripheral nerve sheath tumor (MPNST) is an uncommon neoplasm. Rarely, MPNST may display focal mesenchymal differentiation and this is more frequently encountered in high than low grade lesions. Here we present an example of a low grade MPNST with osteoid, cartilaginous and probably smooth muscle components occurring in the subtemporal fossa of a 26-year-old male patient with no associated neurofibromatosis type 1. The tumor exhibited diffuse S-100 protein expression, whereas immunostainings for epithelial membrane antigen and smooth muscle actin were positive in a portion of neoplastic cells.

Inflammatory Myofibroblastic Tumor of Spinal Canal: Brief Case Report.

We describe a case of an intradural extramedullary inflammatory myofibroblastic tumor of the cervical spine. A 56-year-old woman presented with progressive neck pain, radiating to the right scapula, without any neurologic deficit. Magnetic resonance imaging showed an intradural extramedullary tumor with a dural tail sign, located at the C3-T1 segment with homogeneous contrast enhancement. The patient was operated on for a suspected meningioma. Pathologic examination showed fibrosis and inflammation with infiltration of B and T lymphocytes accompanied by plasmocytes, macrophages, and myofibroblast oocytes. We present the clinical course and review of the literature.

Recurrent Pineocytomalike Papillary Tumor of The Pineal Region: A Case Report and Literature Review.

BACKGROUND: Papillary tumors of the pineal region (PTPRs) are malignant World Health Organization grade II/III tumors; however, they may perfectly mimic benign tumors (e.g., pineocytomas [World Health Organization grade I]). CASE DESCRIPTION: We present a case of a 28-year-old man with a 35-mm tumor of the pineal region. Considering the typical radiological and pathologic presentation, the tumor was first diagnosed as pineocytoma. However, despite first total resection, the tumor recurred after 7 years. The recurrent neoplasm was composed mainly of papillary structures with low-grade atypical cells positive for CKAE1/AE3 and CK18. This categorization led to the final diagnosis of PTPR. The patient underwent adjuvant radiotherapy, which vastly improved his neurologic condition and resulted in significant tumor regression. CONCLUSIONS: This case exemplifies that PTPRs can perfectly mimic pineocytomas and simple staining for cytokeratins may warrant correct diagnosis and better treatment.

Morphological analysis of vascular density in ependymomas.

Ependymomas generally show slow growth rate and are associated with a long clinical history. In some cases however the biology of these tumours is considered to be unpredictable on the basis of histologic criteria. Density of microvessels was shown to serve in various malignant neoplasms as a prognostic factor that correlates with increased risk of metastasis and overall free survival. Some data suggest that density of blood vessels may be of prognostic value also in patients with neuroepithelial tumours. The aim of this study was to determinate whether that observation can be applied to ependymomas. The materials included 51 ependymomas G2 and G3 according to the WHO classification. Vasculature was visualized immunohistochemically in paraffin-embedded sections of tumour samples with CD31 and FVIII antibody. Density of blood vessels was calculated using a computed image analyzing system. The data were statistically evaluated. The density of blood vessels in anaplastic (WHO G3) ependymomas was shown to be significantly higher than that in WHO G2 type of the tumour, while there was no statistical difference between subtypes of WHO G2 ependymomas. The results suggest a connection between density of vasculature and the degree of histological malignancy in gliomas of ependymal derivation.

Ultrastructural heterogeneity of gangliogliomas.

Gangliogliomas are rare brain tumors, composed of neuronal and glial cells mixed in a different proportion. The basic histopathological pattern of gangliogliomas is well recognized but the variable microscopic appearance still can pose a challenge to the neuropathologist. The authors reanalyzed their series of gangliogliomas in the files of two departments of neuropathology. All analyzed tumors fulfilled the WHO histological criteria of ganglioglioma. Seven tumors were examined by electron microscopy. The following ultrastructural features were graded: presence of dense-cored vesicles, synaptic vesicles, synapses and intermediate filaments, abundant basal membranes, dystrophic neurites, autophagic vacuoles, and multivesicular bodies. Most of the neoplastic neurons were large, polyglonal or oval with well-developed subcellular organelles, round nuclei, and prominent nucleoli. In most cases there were abundant dense core vesicles, observed in both the tumor cell bodies as well as in their processes. Synapses were typically observed. Intermediate filaments were abundant in all tumors. The most intriguing ultrastructural finding was abundant presence of autophagic vacuoles. In 4 cases, multivesicular bodies were observed. All of the tumors with multivesicular bodies also contained abundant autophagic vacuoles.

Gliomas: association of histology and molecular genetic analysis of chromosomes 1p, 10q, and 19q.

The aim of our study was to evaluate the frequencies of loss of heterozygosity (LOH) on chromosomes 1p, 10q, and 19q in gliomas and to correlate them with the histological diagnosis and with patient age and gender. We found deletions within chromosome 1p to be significantly associated with the histological subtype of glial tumor (P < 0.05); frequency of 1p deletions increased from astrocytoma (0%) through glioblastoma (31%) and oligoastrocytoma (57%) to oligodendroglioma (63%). In patients with 1p LOH, the odds for having astrocytoma or glioblastoma were approximately 10-fold and 4-fold lower, respectively, than oligodendroglioma. The odds for having oligoastrocytoma were similar to oligodendroglioma (OR = 1.3). The frequency of 10q LOH in patients with glioblastoma was significantly higher than in patients with oligodendroglioma (89% vs. 36%; P < 0.005). In patients with oligodendroglioma, most cases with LOH on chromosome 1p also had LOH 19q (90%), one case of 1p LOH also had a deletion on 10q. Statistical analyses revealed a significant association between deletions on 1p and 19q (P < 0.05). Our data provide evidence that use of molecular genetic analyses of chromosomes 1p, 19q, and 10q might improve the diagnosis of gliomas.

Histological changes of the acetabular labrum in coxarthrosis: labral degeneration and repair.

INTRODUCTION: The current study was designed to describe types of histological changes within the acetabular labrum in the advanced stages of coxarthrosis, in patients requiring total hip arthroplasty (THA). METHODS: 77 consecutive patients without systemic disorders or prior hip surgery, scheduled for THA with 3 types of coxarthrosis: avascular necrosis (AVN), idiopathic, and dysplastic coxarthrosis were analysed. Patient's data: age, gender, side of the involvement, duration of the symptoms were recorded, and standard anteroposterior (AP) radiographic views of the pelvis were obtained. During THA procedure the acetabular labrum was harvested and examined histologically and immunohistochemically. The mean chondrocytes number and density were calculated using morphometric techniques. RESULTS: In 77 analysed acetabular labra the following histological changes were found: heterogenic matrix, foci of granular matrix breakdown, pseudocysts, intralabral c alcifications, chondrocyte apoptosis, inflammatory reaction with lymphocytes infiltration and macrophages infiltration and vascular proliferation with 2 stages of maturation: endothelial cell formation and fully formed blood vessels. The average chondrocytes density was 478 cells in 1 mm2 and significantly decreased with age. CONCLUSIONS: The acetabular labrum is an important part of the process of degeneration of the hip in osteoarthritis (OA). Vascular formation and cellular infiltration found in the damaged fibrocartilage may represent a labral response to degenerative changes.

Three cases of ectopic sphenoid sinus pituitary adenoma.

Introduction: Ectopic sphenoid sinus pituitary adenoma is a rare tumour originating from embryologic remnants of Rathke's pouch. Although it is considered a clinically benign neoplasm, necrosis is encountered in 25% of cases and it can invade adjacent bone structures. Aims: To establish clinical, radiological and histopathological features of ectopic sphenoid sinus pituitary adenoma. Material and methods: Analysis of three cases: two females and one man, aged 61-70. Results: One patient presented with a unilateral hearing loss, the other two with headache and vertigo. They all suffered from type 2 diabetes mellitus. Neurological examination revealed no abnormality. Radiological imaging showed a sphenoid sinus space-occupying soft-tissue lesion with bone erosion in 2 cases and empty sella in 2 patients whereas one had a normal pituitary gland. All were operated on via the transnasal approach. Total resection was achieved in one patient and subtotal in two; in two cases we observed intact sellar dura and in one intact sellar floor. Histopathology showed immunoreactivity for synaptophysin in all cases and cytokeratin in two. The Ki-67 index was less than 2%. Immunohistochemical staining demonstrated growth hormone cells in all cases whereas prolactin and adrenocorticotropin in two. The patients were discharged home in good condition with no neurological deficits. Conclusions: Ectopic sphenoid sinus pituitary adenoma should always be considered in differential diagnosis of sphenoid sinus lesion in the elderly, especially in coexistence with empty sella or type 2 diabetes mellitus. Since ectopic sphenoid sinus pituitary adenoma is a benign lesion, surgical removal is an effective treatment. .

Glioblastoma-derived cells in vitro unveil the spectrum of drug resistance capability - comparative study of tumour chemosensitivity in different culture systems.

Resistance to cancer drugs is a complex phenomenon which could be influenced by in vitro conditions. However, tumour-derived cell cultures are routinely used for studies related to mechanisms of drug responsiveness or the search for new therapeutic approaches. The purpose of our work was to identify the potential differences in drug resistance and response to treatment of glioblastoma with the use of three in vitro models: traditional adherent culture, serum-free spheroid culture and novel adherent serum-free culture.The experimental models were evaluated according to 'stemness state' and epithelial-to-mesenchymal transition (EMT) status, invasion capability and their expression pattern of genes related to the phenomenon of tumour drug resistance. Additionally, the response to drug treatments of three different culture models was compared with regard to the type of cell death.Multi-gene expression profiling revealed differences between examined culture types with regard to the expression pattern of the selected genes. Functionally, the examined genes were related to drug resistance and metabolism, DNA damage and repair and cell cycle control, and included potential therapeutic targets.Cytotoxicity analyses confirmed that environmental factors can influence not only the molecular background of glioblastoma drug-resistance and efficiency of treatment, but also the mechanisms/pathways of cell death, which was reflected by a distinct intensification of apoptosis and autophagy observed in particular culture models. Our results suggest that parallel exploitation of different in vitro experimental models can be used to reveal the spectrum of cancer cell resistance capability, especially regarding intra-heterogeneous glioblastomas.

KCTD11 expression in medulloblastoma is lower than in adult cerebellum and higher than in neural stem cells.

Medulloblastoma (MB) is the most common malignant brain tumor of childhood, and the most frequent associated genetic alteration is loss of heterozygosity on chromosome region 7p13. Two genes mapping to this region, KCTD11 (alias REN) and HIC1, have been proposed as involved in MB pathogenesis. We used real-time polymerase chain reaction in 20 tissue samples of primary MB to examine the transcriptional level of the two genes, with reference to two types of controls: adult cerebellum and fetal neural stem cells. A significant reduction of KCTD11 expression relative to adult normal cerebellum was detected in 14 of 20 (70%) of MB samples. Neural stem cells had even lower levels of KCTD11 expression than did MB. HIC1 gene expression was low ( approximately 100 times lower than KCTD11 expression) in MB, and low also in both adult cerebellum and neural stem cells. Hypermethylation of the 5'UTR or the central region of HIC1 (or both) was detected in a significant number of MB samples, as well as in cerebellum and neural stem cells. Our data suggest that KCTD11 may play an important role in MB tumorigenesis, but do not support the role of HIC1 in this tumor development. We argue that recognition of the gene or genes important in MB tumorigenesis depends in part on defining an appropriate control.

Immunohistochemical expression of progesterone and estrogen receptors in meningiomas.

In this study we present results of investigations of progesterone and estrogen receptors in most frequent, WHO grade I histological types of meningiomas (meningothelial, fibrous, and transitional) and in atypical--WHO grade II variant of these tumors. Samples from 64 tumors were examined. The cohort consisted of 46 WHO grade I (21 transitional, 13 fibrous and 12 meningothelial histologic variants) and of 18 atypical meningiomas. Apart from immunohistochemical examination of progesterone and estrogen receptors, MIB 1 labeling index was estimated. Positive immunoreaction for progesterone receptors was found in 100% meningothelial, 95% transitional, 46% fibrous and 78% atypical variant of meningiomas. Intensity of immunoreaction was stronger in grade I than in grade II tumors. Immunoexpression of estrogen receptors was found in 48% of the investigated meningiomas. No correlation was stated between WHO grade I and grade II tumors, and between meningothelial, transitional and fibrous variants of the neoplasms.