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.

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.

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.

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. .

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.

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.

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).

TP53 promoter methylation in primary glioblastoma: relationship with TP53 mRNA and protein expression and mutation status.

Reduced expression of TP53 by promoter methylation has been reported in several neoplasms. It remains unclear whether TP53 promoter methylation is associated with reduced transcriptional and protein expression in glioblastoma (GB). The aim of our work was to study the impact of TP53 methylation and mutations on TP53 mRNA level and protein expression in 42 molecularly characterized primary GB tumors. We also evaluate the impact of all molecular alterations on the overall patient survival. The frequency of TP53 promoter methylation was found in 21.4%. To the best of our knowledge, this is the first report showing such high frequency of TP53 promoter methylation in primary GB. There was no relation between TP53 promoter methylation and TP53 mRNA level (p=0.5722) and between TP53 promoter methylation and TP53 protein expression (p=0.2045). No significant associations were found between TP53 mRNA expression and mutation of TP53 gene (p=0.9076). However, significant association between TP53 mutation and TP53 protein expression was found (p=0.0016). Our data suggest that in primary GB TP53 promoter methylation does not play a role in silencing of TP53 transcriptional and protein expression and is probably regulated by other genetic and epigenetic mechanisms associated with genes involved in the TP53 pathway.

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.

Screening for EGFR amplifications with a novel method and their significance for the outcome of glioblastoma patients.

Glioblastoma is a highly aggressive tumour of the central nervous system, characterised by poor prognosis irrespective of the applied treatment. The aim of our study was to analyse whether the molecular markers of glioblastoma (i.e. TP53 and IDH1 mutations, CDKN2A deletion, EGFR amplification, chromosome 7 polysomy and EGFRvIII expression) could be associated with distinct prognosis and/or response to the therapy. Moreover, we describe a method which allows for a reliable, as well as time- and cost-effective, screening for EGFR amplification and chromosome 7 polysomy with quantitative Real-Time PCR at DNA level. In the clinical data, only the patient's age had prognostic significance (continuous: HR = 1.04; p<0.01). At the molecular level, EGFRvIII expression was associated with a better prognosis (HR = 0.37; p = 0.04). Intriguingly, EGFR amplification was associated with a worse outcome in younger patients (HR = 3.75; p<0.01) and in patients treated with radiotherapy (HR = 2.71; p = 0.03). We did not observe any difference between the patients with the amplification treated with radiotherapy and the patients without such a treatment. Next, EGFR amplification was related to a better prognosis in combination with the homozygous CDKN2A deletion (HR = 0.12; p = 0.01), but to a poorer prognosis in combination with chromosome 7 polysomy (HR = 14.88; p = 0.01). Importantly, the results emphasise the necessity to distinguish both mechanisms of the increased EGFR gene copy number (amplification and polysomy). To conclude, although the data presented here require validation in different groups of patients, they strongly advocate the consideration of the patient's tumour molecular characteristics in the selection of the therapy.

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.

Reduced expression of ELAVL4 in male meningioma patients.

Meningioma is a frequently occurring tumor of the central nervous system. Among many genetic alternations, the loss of the short arm of chromosome 1 is the second most frequent chromosomal abnormality observed in these tumors. Here, we focused on the previously described and well-established minimal deletion regions of chromosome 1. In accordance with the Knudson suppressor theory, we designed an analysis of putative suppressor genes localized in the described minimal deletion regions. The purpose was to determine the molecular background of the gender-specific occurrence of meningiomas. A total of 149 samples were examined for loss of heterozygosity (LOH). In addition, 57 tumor samples were analyzed using real-time polymerase chain reaction. We examined the association between the expression of selected genes and patient age, gender, tumor grade and presence of 1p loss. Furthermore, we performed an analysis of the most stable internal control for real-time analysis in meningiomas. LOH analysis revealed gender-specific discrepancies in the frequency of 1p aberrations. Moreover, statistical correlation between the gene expression level and gender was significant for the ELAVL4 gene as we found it to be lower in males than in females. We conclude that meningiomas present different features depending on patient gender. We suggest that ELAVL4 can be involved in the pathogenesis of meningiomas in male patients.

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.

Extracerebral metastases of glioblastoma have a different vasculature than primary tumour. A case report of glioblastoma extracranial metastases.

The paper presents a case report of a 38-year-old female suffering from metastatic glioblastoma in the jugular lymph node that developed 9 months after craniotomy and tumorectomy in the left temporal region of the brain. The histological evaluation of metastatic tumour reveals lower density of vasculature as well as less significant pathologic changes in blood vessels morphology in comparison to primary tumour. Moreover, in this report we present cytological characteristics of the material obtained by fine needle aspiration of the metastatic mass.

[Analysis of HIF-1α and COX-2 expression in tumor stroma and correlation with the degree of neoplasm invasiveness in laryngeal cancer--preliminary study]. / Analiza ekspresji HIF-1α i COX-2 w utkaniu guza oraz korelacja ze stopniem inwazyjnosci zmian nowotworowych u chorych z rakiem krtani--badania wstepne.

INTRODUCTION: Despite extensive research in the field of molecular biology, immunology and histopathology, prognostically unambiguous morphological indicators of the invasiveness of tumor which allow the prediction of disease course in laryngeal cancer have not yet been identified. The aim of this study was to analyze gene and protein expression of HIF-1a and COX-2 in the tumor stroma and to find relationships between clinical and morphological features (pT, G, depth of tumor invasion, plasmalymphocytic infiltration) and certain markers in squamous cell laryngeal carcinoma. MATERIAL AND METHODS: We analyzed a group of 59 patients with verified squamous cell carcinoma of the larynx. The pathological evaluation was included pTNM classification criteria, depth of invasion according to tumor front grading and degree of morphological differentiation. Quantitative analysis of the amplified product in real time (real-time RT2-PCR) for estimation of mRNA HIF-1a and COX-2 expression in tumor cells were used. The level of HIF-1a and COX-2 protein expression by Western blot analysis was determined. RESULTS: In studied group of laryngeal cancers significant differences in expression of certain molecules analyzed in tumor tissue and noncancerous epithelium of the larynx have been shown. Increased expression of HIF-1a and COX-2 at both mRNA and protein level was indicator of greater aggressiveness of the tumor, evaluated on the basis of clinical and morphological features in laryngeal squamous cell carcinomas. Tumors with the most invasive growth (pT4 stage, low-differentiated tumors G3, neoplastic infiltration with invasion of cartilage of the larynx) had the highest expression of HIF-1a and COX-2 at the mRNA and protein level. CONCLUSIONS: The study pointed out the direction for further research to find unambiguous indicators for estimation of tumor invasiveness and the possibility of practical use of HIF-1a and COX-2 mRNA and protein level assessment as important methods for determining the advancement of clinical and morphological changes in laryngeal cancer, thereby selecting an appropriate model of treatment.

[Expression of Foxp3 and RORgamma t in peripheral blood mononuclear cells in patients with laryngeal carcinoma as indicators of tumor stage--preliminary study]. / Ocena ekspresji Foxp3 i RORgamma t w jednojadrzastych komórkach krwi obwodowej u chorych z rakiem krtani jako wskazników zaawansowania zmian nowotworowych--badania wstepne.

INTRODUCTION: The degree of activation of cells involved in cellular immune response against tumor antigens (cytotoxic lymphocytes Tc) as well as efficiency of the mechanisms which promote immunosuppression (Treg - regulatory cells CD4(+)CD25(+)Foxp3(+)) may determine the course of the neoplastic disease. The aim of this study was to assess the function of autologous peripheral blood mononuclear cells (PBMCs) involved in the immunological processes on the basis of expression of Foxp3 and RORgamma t molecules as well as analysis of the relationships with clinical and morphological features of the tumor (pT and pN stage, G feature, degree of invasiveness according to the TFG classification) in laryngeal carcinoma. MATERIAL AND METHODS: The analysis included a group of 59 patients with verified squamous cell carcinoma of the larynx. In the pathologic evaluation pTNM classification criteria, depth of invasion and degree of histological differentiation were used. Expression levels of mRNA for Foxp3 and RORgamma t in peripheral blood mononuclear cells by quantitative analysis of the amplified product in real time (real-time RT(2)-PCR) were evaluated. The level of Foxp3 and RORgamma t protein expression by Western blot analysis was determined. RESULTS: In squamous cell carcinomas of the larynx, with the highest tumor aggressiveness the significantly highest level of mRNA and protein expression for Foxp3 molecule were observed. The severity of Foxp3 expression at both gene and protein level were positively linearly correlated with the degree of local extent of the tumor (pT3-4), depth of invasion (invasion of cartilage) and the degree of histological differentiation (low-differentiated tumors G3). In the study group of laryngeal cancers significantly lower level of RORgamma t expression in carcinomas with less invasive changes (pT1-2, high-differentiated tumors G1, carcinomas with microinvasion without evidence of invasion beyond the lamina propria) was also noted. CONCLUSIONS: The study results indicate the important role of immune cell activity as indicators of advancement of clinical and morphological changes in squamous cell carcinoma of the larynx.

Analysis of the prognostic significance of selected morphological and immunohistochemical markers in ependymomas, with literature review.

AIM: Ependymal tumours are relatively uncommon primary neoplasms of the central nervous system. Histological criteria distinguishing ependymoma and anaplastic ependymoma are not clear-cut and other parameters are required to allow more precise prognostication in these tumours. We analysed the histological and immunohistochemical features of these tumours (Ki-67, cyclin D1, EGFR, hTERT, Olig2) and correlated them with the clinical outcome. MATERIAL AND METHODS: We analysed 39 patients with grade II ependymoma (30) and anaplastic ependymoma (9). Twenty-eight tumours developed in children and the remaining 11 patients were adults with intracranial and intraspinal tumours. Eighteen patients died during the follow-up period. RESULTS AND CONCLUSIONS: Overall survival was reduced significantly for paediatric patients and patients with intracranial tumour. High-grade tumours, increased mitotic index and increased cellularity had an unfavourable influence on survival. Other histological parameters such as nuclear atypia, necrosis and microvascular proliferation did not alter the survival rate. Increased Ki-67 and cyclin D1 indices correlated with worse prognosis. Furthermore, any level of cyclin D1 expression in WHO grade II ependymomas was strongly associated with higher risk of death. No correlation was identified between Olig2, EGFR and hTERT expression and the outcome of the patients.