Targeted sequencing of cancer-related genes reveals a recurrent TOP2A variant which affects DNA binding and coincides with global transcriptional changes in glioblastoma.

High-grade gliomas are aggressive, deadly primary brain tumors. Median survival of patients with glioblastoma (GBM, WHO grade 4) is 14 months and <10% of patients survive 2 years. Despite improved surgical strategies and forceful radiotherapy and chemotherapy, the prognosis of GBM patients is poor and did not improve over decades. We performed targeted next-generation sequencing with a custom panel of 664 cancer- and epigenetics-related genes, and searched for somatic and germline variants in 180 gliomas of different WHO grades. Herein, we focus on 135 GBM IDH-wild type samples. In parallel, mRNA sequencing was accomplished to detect transcriptomic abnormalities. We present the genomic alterations in high-grade gliomas and the associated transcriptomic patterns. Computational analyses and biochemical assays showed the influence of TOP2A variants on enzyme activities. In 4/135 IDH-wild type GBMs we found a novel, recurrent mutation in the TOP2A gene encoding topoisomerase 2A (allele frequency [AF] = 0.03, 4/135 samples). Biochemical assays with recombinant, wild type (WT) and variant proteins demonstrated stronger DNA binding and relaxation activity of the variant protein. GBM patients carrying the altered TOP2A had shorter overall survival (median OS 150 vs 500 days, P = .0018). In the GBMs with the TOP2A variant we found transcriptomic alterations consistent with splicing dysregulation. luA novel, recurrent TOP2A mutation, which was found exclusively in four GBMs, results in the TOP2A E948Q variant with altered DNA binding and relaxation activities. The deleterious TOP2A mutation resulting in transcription deregulation in GBMs may contribute to disease pathology.

Exploring Novel Therapeutic Opportunities for Glioblastoma Using Patient-Derived Cell Cultures.

Glioblastomas (GBM) are the most common, primary brain tumors in adults. Despite advances in neurosurgery and radio- and chemotherapy, the median survival of GBM patients is 15 months. Recent large-scale genomic, transcriptomic and epigenetic analyses have shown the cellular and molecular heterogeneity of GBMs, which hampers the outcomes of standard therapies. We have established 13 GBM-derived cell cultures from fresh tumor specimens and characterized them molecularly using RNA-seq, immunoblotting and immunocytochemistry. Evaluation of proneural (OLIG2, IDH1R132H, TP53 and PDGFRα), classical (EGFR) and mesenchymal markers (CHI3L1/YKL40, CD44 and phospho-STAT3), and the expression of pluripotency (SOX2, OLIG2, NESTIN) and differentiation (GFAP, MAP2, ß-Tubulin III) markers revealed the striking intertumor heterogeneity of primary GBM cell cultures. Upregulated expression of VIMENTIN, N-CADHERIN and CD44 at the mRNA/protein levels suggested increased epithelial-to-mesenchymal transition (EMT) in most studied cell cultures. The effects of temozolomide (TMZ) or doxorubicin (DOX) were tested in three GBM-derived cell cultures with different methylation status of the MGMT promoter. Amongst TMZ- or DOX-treated cultures, the strongest accumulation of the apoptotic markers caspase 7 and PARP were found in WG4 cells with methylated MGMT, suggesting that its methylation status predicts vulnerability to both drugs. As many GBM-derived cells showed high EGFR levels, we tested the effects of AG1478, an EGFR inhibitor, on downstream signaling pathways. AG1478 caused decreased levels of phospho-STAT3, and thus inhibition of active STAT3 augmented antitumor effects of DOX and TMZ in cells with methylated and intermediate status of MGMT. Altogether, our findings show that GBM-derived cell cultures mimic the considerable tumor heterogeneity, and that identifying patient-specific signaling vulnerabilities can assist in overcoming therapy resistance, by providing personalized combinatorial treatment recommendations.

Gliosarcoma Is Driven by Alterations in PI3K/Akt, RAS/MAPK Pathways and Characterized by Collagen Gene Expression Signature.

Gliosarcoma is a very rare brain tumor reported to be a variant of glioblastoma (GBM), IDH-wildtype. While differences in molecular and histological features between gliosarcoma and GBM were reported, detailed information on the genetic background of this tumor is lacking. We intend to fill in this knowledge gap by the complex analysis of somatic mutations, indels, copy number variations, translocations and gene expression patterns in gliosarcomas. Using next generation sequencing, we determined somatic mutations, copy number variations (CNVs) and translocations in 10 gliosarcomas. Six tumors have been further subjected to RNA sequencing analysis and gene expression patterns have been compared to those of GBMs. We demonstrate that gliosarcoma bears somatic alterations in gene coding for PI3K/Akt (PTEN, PI3K) and RAS/MAPK (NF1, BRAF) signaling pathways that are crucial for tumor growth. Interestingly, the frequency of PTEN alterations in gliosarcomas was much higher than in GBMs. Aberrations of PTEN were the most frequent and occurred in 70% of samples. We identified genes differentially expressed in gliosarcoma compared to GBM (including collagen signature) and confirmed a difference in the protein level by immunohistochemistry. We found several novel translocations (including translocations in the RABGEF1 gene) creating potentially unfavorable combinations. Collected results on genetic alterations and transcriptomic profiles offer new insights into gliosarcoma pathobiology, highlight differences in gliosarcoma and GBM genetic backgrounds and point out to distinct molecular cues for targeted treatment.

Cytokines in the pathogenesis of hemophilic arthropathy.

Hemophilic arthropathy (HA) is one of the most common and typical manifestation in the course of recurrent bleeding episodes in patients with hemophilia. Clinical and subclinical joint bleeding episodes gradually lead to irreversible changes manifesting themselves as pain, progressing ankylosis, marked limitation of the range of motion, muscle atrophy and osteoporosis commonly concomitant with joint deformity resulting from chronic proliferative synovitis and both cartilage and bone degeneration leading to the final functional impairment of the joint. In spite of numerous studies, the pathophysiology of HA has not been fully elucidated, especially as regards immunopathological mechanisms which are associated with the subclinical and early stage of the disease and to be more precise, with chronic joint inflammation. It needs to be emphasized that the pathophysiological processes occurring in a joint with HA are most probably highly mediated by interactions within the cytokine network and other inflammatory mediators present in the tissues of affected joint. Among numerous compounds participating in the induction of an inflammatory process in the pathogenesis of HA, cytokines seem to play a leading role. The most important group controlling the disease seems to be well known inflammatory cytokines, including IL-1ß, TNFα and IL-6. The second group with antagonistic effect is formed by anti-inflammatory cytokines such as IL-4 and IL-10. The role of inflammatory and anti-inflammatory cytokines in the pathogenesis of HA with respect to cellular and intracellular signaling pathways is still under investigation. This review, summarizes and discusses the current knowledge about cytokine network in the pathogenesis of HA, indicating possible molecular and cellular mechanisms that may provide potential new therapeutic directions.

Down-regulation of IKKß expression in glioma-infiltrating microglia/macrophages is associated with defective inflammatory/immune gene responses in glioblastoma.

Glioblastoma (GBM) is an aggressive malignancy associated with profound host immunosuppression. Microglia and macrophages infiltrating GBM acquire the pro-tumorigenic, M2 phenotype and support tumor invasion, proliferation, survival, angiogenesis and block immune responses both locally and systematically. Mechanisms responsible for immunological deficits in GBM patients are poorly understood. We analyzed immune/inflammatory gene expression in five datasets of low and high grade gliomas, and performed Gene Ontology and signaling pathway analyses to identify defective transcriptional responses. The expression of many immune/inflammatory response and TLR signaling pathway genes was reduced in high grade gliomas compared to low grade gliomas. In particular, we found the reduced expression of the IKBKB, a gene coding for IKKß, which phosphorylates IκB proteins and represents a convergence point for most signal transduction pathways leading to NFκB activation. The reduced IKBKB expression and IKKß levels in GBM tissues were demonstrated by qPCR, Western blotting and immunohistochemistry. The IKKß expression was down-regulated in microglia/macrophages infiltrating glioblastoma. NFκB activation, prominent in microglia/macrophages infiltrating low grade gliomas, was reduced in microglia/macrophages in glioblastoma tissues. Down-regulation of IKBKB expression and NFκB signaling in microglia/macrophages infiltrating glioblastoma correlates with defective expression of immune/inflammatory genes and M2 polarization that may result in the global impairment of anti-tumor immune responses in glioblastoma.

Thyroid hormones in the central nervous system (CNS) and their effect on neoplasm formation, particularly on the development and course of glioblastoma multiforme - research hypothesis.

The present study is aimed to present the potential role of thyroid hormones (TH) in the pathogenesis of glioblastoma multiforme (GBM). In first part of this presentation the effect of general homeostasis of TH on GBM formation and course was shown. Then the evidence concerning present state of the knowledge about active transport of TH to the brain, the role of iodothyronine deiodinase type 2 and 3 in the setting concentration of T3 in the brain and GBM cells, and finally knowledge about the role of genomic (TH nuclear receptors THRA and THRB) and non-genomic modes (membrane integrin receptor αvß3) of action of TH and its importance for GBM was outlined. The last part of this presentation was devoted to generally approved signalling pathways leading to the formation and the clinical course of GBM, showing at the same time evidence that each of the pathways is affected by particular TH actions. In conclusion it is suggested that TH is one of the pathogenetic factors for GBM and as such can have practical implications for the formation and course and treatment of this tumour.

Morphometric predictors of posterior circulation aneurysms risk rupture.

OBJECTIVES: Intracranial aneurysms (IAs) of the posterior circulation (PC) rupture more frequently and their morbidity and mortality rates are higher compared to anterior circulation. Morphological parameters such as size ratio (SR), inflow angle and parent artery geometry are believed to contribute significantly in determining IA risk rupture. The aim of this study is to establish angiography-based morphometric predictors of PC IA risk rupture. METHODS: A retrospective analysis of 58 patients with PC aneurysms was incorporated into the study. The following independent variables were measured: aneurysm dome size, neck size, parent artery size, SR, neck to parent artery ratio, and inflow angle. All aneurysms were divided into ruptured and unruptured groups. The stepwise logistic regression analysis was applied to establish the predictors of PC aneurysm risk rupture. RESULTS: 58 patients with 27 unruptured and 31 ruptured PC cerebral aneurysms were analyzed. The mean aneurysm dome, neck and parent vessel diameters were 8·49±3·5 mm, 2·46±1·4 mm, and 3·92±1·6 mm, respectively. Size ratio was 2·26±0·6; dome/neck 3·45±0·8 and inflow angle 115·2±22°. The relevant difference between unruptured and ruptured groups was: SR (1·91 vs 2·48), aneurysm dome (7·96 vs 8·95 mm), dome/neck (3·77 vs 3·18), and inflow angle (103·7 vs 125·2°). The significant predictive value was reached for inflow angle (OR 1·05; Cl 95% 1·01 to 1·1) and SR (OR 3·53; Cl 95% 1·09 to 11·5). Cut-off value on receiver operating characteristic curve for inflow angle (113·1°; sensitivity 67·7% and specificity 81·5%) and SR (1·99; sensitivity 77·4% and specificity 63%). DISCUSSION: The SR and inflow angle proved to be relevant predictors in estimating the aneurysm risk rupture of the posterior cerebral circulation.

Quality of life and depressive symptoms in Parkinson's disease after subthalamic deep brain stimulation: a 2-year follow-up study.

AIM: To assess the correlation between quality of life (QoL), depressive symptoms and motor signs in patients with Parkinson disease after subthalamic deep brain stimulation (DBS STN). MATERIAL AND METHODS: 74 patients, average age 55.6 ± 7 and duration of disease 12.3 years ± 3.8, treated with l STN DBS for PD were included in the study. All patients were evaluated with (UPDRS III), (PDQ-39) (BDI) at baseline and at 6, 12, and 24-month follow up. All patients were also stratified into three groups depending on UPDRS III improvement ( < 30%, 30-60%, > 60%). RESULTS: Scores in all scales significantly decreased from baseline. The improvement in PDQ-39 was 43.3%, in BDI 25.3 %; UPDRS-III 55.5% at 6 months. At 24 months, motor results and QoL deteriorated by 15.6% and 19.6% respectively. BDI remained unchanged. Mean scores at baseline in PDQ-39 were group I 67.4 ± 29.7; II 64.8 ± 32.0; III 53.4 ± 22.0 and for BDI, group I 17.4 ± 12.04; II 14.0 ± 9.7; III- 15.1 ± 10.55. Scores decreased significantly with DBS at 6-month follow-up and mean change was: PDQ-39, group I 42.7%, II- 40.7%, III 51.6%; BDI group I 23%, II 28.1%, III 23.3 %. CONCLUSION: Reduction of depressive symptoms, motor signs and improvement of QoL in PD after DBS STN are closely related. Improvement of QoL depends significantly on motor symptoms.

Alternative splicing of iodothyronine deiodinases in pituitary adenomas. Regulation by oncoprotein SF2/ASF.

Pituitary tumors belong to the group of most common neoplasms of the sellar region. Iodothyronine deiodinase types 1 (DIO1) and 2 (DIO2) are enzymes contributing to the levels of locally synthesized T3, a hormone regulating key physiological processes in the pituitary, including its development, cellular proliferation, and hormone secretion. Previous studies revealed that the expression of deiodinases in pituitary tumors is variable and, moreover, there is no correlation between mRNA and protein products of the particular gene, suggesting the potential role of posttranscriptional regulatory mechanisms. In this work we hypothesized that one of such mechanisms could be the alternative splicing. Therefore, we analyzed expression and sequences of DIO1 and DIO2 splicing variants in 30 pituitary adenomas and 9 non-tumorous pituitary samples. DIO2 mRNA was expressed as only two mRNA isoforms. In contrast, nine splice variants of DIO1 were identified. Among them, five were devoid of exon 3. In silico sequence analysis of DIO1 revealed multiple putative binding sites for splicing factor SF2/ASF, of which the top-ranked sites were located in exon 3. Silencing of SF2/ASF in pituitary tumor GH3 cells resulted in change of ratio between DIO1 isoforms with or without exon 3, favoring the expression of variants without exon 3. The expression of SF2/ASF mRNA in pituitary tumors was increased when compared with non-neoplastic control samples. In conclusion, we provide a new mechanism of posttranscriptional regulation of DIO1 and show deregulation of DIO1 expression in pituitary adenoma, possibly resulting from disturbed expression of SF2/ASF.

Pain perception in patients with Parkinson's disease.

Abnormalities in pain perception are a part of the clinical picture in Parkinson's disease (PD) and belong to the category of non-motor symptoms. Two groups of patients were included in this study: (i) an experimental group of 36 patients with PD who were eligible for subthalamic deep brain stimulation (the experimental group [EG]) and (ii) a control group (CG) of 34 patients with a space-occupying lesion who were admitted for a framed stereotactic biopsy. Stereotactic frame fixation was used in both groups as a nociceptive stimulus. All participants were assessed for pain perception with two kinds of visual analogue scales (VAS) (a non-color VAS [ncVAS] and a color VAS [cVAS]) immediately after the stimulus (EG - ncVAS 1 and cVAS 1; CG - ncVAS 3 and cVAS 3) and 24 hours later (EG - ncVAS 2 and cVAS 2; CG - ncVAS 4 and cVAS 4). The means for the two pain scores assessed directly after frame fixation were 3.59 (ncVAS 1) and 3.06 (cVAS 1) for patients in the EG, while the mean ncVAS was 3, and the mean cVAS 3 was 6.1 for those in the CG. The pain intensity was significantly lower for patients with PD (EG) compared to those in the CG for both ncVAS and cVAS (p<0.05 for each measure). The mean pain scores for ncVAS and cVAS measured 24 hours after the procedure were 3.18 and 2.79 for patients with PD (EG) and 6.10 and 5.77 for those in the CG, respectively. Pain intensity measured 24 hours after the procedure was significantly lower in those with PD (EG) compared to the CG. This study has demonstrated that pain perception in patients with PD is significantly lower than pain perception in non-parkinsonian patients.

Endovascular embolization of ruptured and unruptured posterior circulation aneurysms. A multi-factor analysis.

AIM: Complete aneurysm obliteration reduces the risk of rebleeding and is an important goal of the aneurysm treatment. MATERIAL AND METHODS: A retrospective analysis of 63 patients undergoing endovascular treatment of posterior circulation aneurysms. The occlusion rate was stratified to three groups: complete, incomplete and partial. In the analysis compared the influence of the selected characteristics: age, sex, WFNS grade, aneurysm location, size of the aneurysm dome and neck, neck to dome ratio, on the effectiveness of embolization in each group. A multi-factor analysis with probit model and linear regression was applied to assess the impact of all characteristics on the complete occlusion. RESULTS: In the series of 63 single aneurysms, 51 were ruptured and 12 unruptured aneurysms. Complete occlusion was achieved in 36 (57.1%), incomplete in 15 (23.8%), partial in 12 (19%) patients. In patients with the neck size of 1-2 mm the complete occlusion was in 75% (24/32) incomplete in 12,5% (4/32), while when the neck size was 2-4 mm these rates were 38.7% (12/31) and 29% (9/31) respectively. The predictor of total occlusion in probit and linear regression model was only one independent variable, the narrow-neck size. CONCLUSION: Multi-factor analysis found that the strongest predictor of the complete occlusion is the aneurysm neck size.

Microlesion effect as a predictor of the effectiveness of subthalamic deep brain stimulation for Parkinson's disease.

BACKGROUND: Microlesion effect (MLE) is a commonly observed phenomenon after electrode insertion into the subthalamic nucleus (STN) for deep brain stimulation (DBS). OBJECTIVES: The aim of this study was to determine the presence of the MLE in the early postoperative period and the relationship between MLE and STN DBS. METHODS: 74 patients with Parkinson's disease were included in this study. Motor symptoms were evaluated preoperatively, within 48 h after electrode implantation and at 6 months with United Parkinson's Disease Rating Scale part III (UPDRS-III). According to the improvement level with MLE, all participants were stratified into three groups: (1) less than 20%; (2) 20-40%, and (3) more than 40% in OFF medication states. The degree of improvement in UPDRS-III with DBS ON for each MLE group was assessed at the 6-month follow-up. Regression analysis was applied for the evaluation of the relationship between MLE and improvement with DBS ON. RESULTS: Mean results in UPDRS-III with the MLE in ON and OFF medication states were 22.1 ± 10.5 and 42.1 ± 14 points, respectively. At the 6-month follow-up, with active stimulation, results tended to further ameliorate to 14.6 (59.4%) points in ON and 20.8 (55.3%) in OFF. Mean improvement in MLE groups were: 33.6% group 1, 47.5% group 2 and 61.4% group 3. Regression analysis revealed a positive correlation between the MLE and results at 6 months with DBS ON. CONCLUSION: Results proved the presence of MLE in the early postoperative period. Furthermore, a positive correlation between MLE and improvement degree with active stimulation was observed.

Deep brain stimulation for refractory epilepsy.

Deep brain stimulation (DBS) is a method of treatment utilized to control medically refractory epilepsy (RE). Patients with medically refractory epilepsy who do not achieve satisfactory control of seizures with pharmacological treatment or surgical resection of the epileptic focus and those who do not qualify for surgery could benefit from DBS. The most frequently used stereotactic targets for DBS are the anterior thalamic nucleus, subthalamic nucleus, central-medial thalamic nucleus, hippocampus, amygdala and cerebellum. The DBS is believed to be an effective method of treatment for various types of epilepsy among adults and adolescents. Side effects may be associated with implantation of electrodes and with the stimulation itself. An increasing number of publications and growing interest in DBS application for RE may result in standardization of the qualification and treatment protocol for RE with DBS.

Motor cortex stimulation in the treatment of neuropathic pain.

BACKGROUND AND PURPOSE: Despite the rapid development of neuropharmacotherapy, medical treatment of neuropathic pain (NP) still constitutes a significant socioeconomic problem. The authors herein present a group of patients treated with motor cortex stimulation (MCS) for NP of various types and aetiologies. MATERIAL AND METHODS: Our cohort included 12 female and 11 male NP patients aged 53 ± 16 treated with MCS. Eleven patients were diagnosed with neuropathic facial pain (NFP), 8 with hemi-body neuropathic pain (HNP), and 4 with deafferentation pain (DP). Prior to surgery, 16 out of 23 patients were treated with repetitive transcranial magnetic stimulation (rTMS), with a positive response in 10 cases. Pain intensity in our group was evaluated with the visual analogue scale (VAS) one month before and three months after MCS implantation. RESULTS: Improvement on the VAS was reported in the whole group of patients (p < 0.001). The best results were reported in the NFP group (p < 0.001) while the worst ones were noted in the DP group (p = 0.04). Anamnesis duration positively correlated with outcome. Infection forced the authors to permanently remove the system in one case. There were no other complications in the group. CONCLUSIONS: Minimally invasive, safe neuromodulative treatment with MCS permits neuropathic pain control with good efficacy. The type of neuropathic pain might be a prognostic factor.

Statistical analysis of the intracranial procedures in Poland in 2008-2009.

BACKGROUND AND PURPOSE: Quantitative and qualitative analysis of neurosurgical procedures provides important data for assessment of the development and trends in the field of neurosurgery. The authors present statistical data on intracranial procedures (IPs) performed in Poland in 2008-2009. MATERIAL AND METHODS: Data on IPs come from reports of the National Health Fund, grouped according to the system of Diagnosis-Related Groups, group A - nervous system diseases. Data concerning the year 2009 include all IPs performed in Poland. Data from the second half of 2008 to 2009 (18 months) come from 35 neurosurgical centers in Poland, divided by provinces. We analyzed the number of IPs, the cost of procedures, duration of hospitalization and deaths. RESULTS: 20 849 IPs were performed in Poland in 2009. The most common procedure was A12 (6807; 32.65%), and the rarest was A04 (96; 0.46%). The annual cost of all IPs was 228 599 956 PLN. Average cost of the procedure ranged from 1578 PLN (A14) to 47 940 PLN (A03). Duration of the hospitalization ranged between 3 days (A14) and 12 days (A12). The highest percentage of deaths was reported for A01 (n = 1050, 19.06%). Reports from 35 neurosurgical centers in the second half of 2008 and 2009 showed the highest number of IPs per 100 000 population in Kujawsko-Pomorskie (93) and the lowest in Wielkopolskie (27) and Podkarpackie (27). The highest number of IPs (1669) was performed in neurosurgical center M1 (Malopolskie), and the lowest (99) in W1 (Wielkopolskie). CONCLUSIONS: A significant disparity in the number of IPs performed in different centers in Poland was observed.

Cerebral amyloid angiopathy manifested as a brain tumour. Clinical and neuropathological characteristics of two cases.

We present two cases (female and male patients, aged 64 and 38, respectively) of focal mass lesions mimicking a brain tumour: one with cognitive function deficit, memory troubles, behavioral changes and left hemiparesis, the other with difficulty in orientation and right hemiparesis. General physical and neurological examinations, laboratory tests and neuroimaging were used to diagnose the cases. Both of them showed nonspecific changes in the brain tissue and the brain tumour was suspected. In the first case MRI scan revealed two pathological masses in the right frontal region and hemorrhagical focus with destructions inside lesions. Second patient's MRI scan revealed a pathological mass at the interface of the left temporal and occipital regions. The neurosurgical procedure was performed. The final diagnosis was established on the basis of neuropathological examination of postoperative material. On light microscopy examination a severe cerebral amyloid angiopathy (CAA) was revealed. Amyloidoma was excluded due to the absence of amorphous material and eosynophylic masses. Tumefactive CAA is a rare condition. These two cases of focal, tumefactive, masslike lesions of diffuse cerebral amyloid angiopathy are reported because of diagnostic dilemmas. In patients with history of memory disfunction, neurological deterioration and different multiple changes observed in CT and MRI scans, such as hemorrhagic infarcts and ischemic cerebral lesions, CAA should be suspected. The imaging findings make a distinction between tumefactive CAA and brain tumours like gliomas difficult. A differential diagnosis of CAA and amyloidoma plays a significant role in a neuropathological examination.

Fluorescence-guided resection of primary and recurrent malignant gliomas with 5-aminolevulinic acid. Preliminary results.

BACKGROUND AND PURPOSE: Extent of resection plays a key role in the treatment of malignant gliomas (MGs). Patients with complete glioma removal, followed by chemoradiation, obtain the longest overall and progression-free survival. Fluorescence-guided resection of MGs enables intraoperative visualization of glioma tissue and increases control of the resection. The authors present preliminary results of 5-aminolevulinic acid (5-ALA) application during the resection of primary and recurrent MGs. MATERIAL AND METHODS: Six patients with either a suspected malignant glioma based on magnetic resonance imaging (MRI) or with recurrent glioblastoma multiforme were enrolled in the study. The extent of resection was calculated according to the postoperative MRI performed within 72 hours. Preoperative and early postoperative neurological status and Karnofsky Performance Scale (KPS) were compared. RESULTS: Fluorescence of tumour tissue was observed in 5/6 patients (five with the histopathological diagnosis of glioblastoma multiforme and one with neurotoxoplasmosis and AIDS). Complete tumour resection was achieved in 5 patients. Postoperative KPS and neurological status deteriorated in 2 cases. Radiotherapy and chemotherapy did not interfere with the sensitivity of the fluorescence guided tumour visualization. CONCLUSIONS: Fluorescence-guided resection of primary and recurrent MGs with 5-ALA improves control of the tumour resection. It enables the cytoreduction to be maximized but experience in neuro-oncological surgery is required to avoid serious, postoperative neurological deficits.

Cocaine self-administration in Warsaw alcohol high-preferring (WHP) and Warsaw alcohol low-preferring (WLP) rats.

Individuals prone to drug self-administration may be vulnerable not only to a single drug reinforcer but to a variety of drug reinforcers. It has been shown that two thirds of alcoholics regularly use drugs other than ethanol (alcohol). Up to 30% of alcohol-dependent patients report concurrent misuse of cocaine. The aim of the present study was to investigate intravenous cocaine self-administration in selectively bred, alcohol-preferring WHP (Warsaw high-preferring) and non-preferring WLP (Warsaw low-preferring) rats. It was hypothesized that WHPs could be more prone to cocaine self-administration in comparison to WLPs. Rats from both lines were allowed to nose-poke for cocaine infusions (0.33 mg/kg/infusion) under the FR-1, FR-2, and FR-3 schedule of reinforcement. Dose-response curves were assessed with increasing doses of cocaine (0.03, 0.1, 0.33, 1.0mg/kg/infusion). The WHP and WLP rats did not differ in cocaine self-administration. Both groups quickly acquired nose-poke responding for cocaine, presented a similar response profile when the schedule of reinforcement was increased from FR-1 to FR-3, and similar sensitivity to cocaine in the dose-response test. The present results may indicate that the selective breeding of alcohol-preferring WHP and alcohol non-preferring WLP rats did not lead to differences in cocaine's rewarding effects as assessed in the self-administration procedure.

Histopathological patterns of papillary tumour of the pineal region.

Papillary tumour of the pineal region (PTPR) is a rare neoplasm that has been formally included in the 2007 WHO classification of central nervous system tumours. The critical diagnosis of this neoplasm is often difficult because of its similarity to other primary or secondary papillary lesions of the pineal region, including parenchymal pineal tumours, papillary ependymoma, papillary meningioma, choroid plexus papilloma and metastatic papillary carcinoma. We present the variability of the histopathological pattern in three cases of PTPR. All cases showed predominant epithelial-like morphology but with various degrees of papillary formation and intensity of cellular pleomorphism. One tumour was highly cystic and exhibited cellular sheets containing vessels covered by several layers of uniform columnar to cuboidal tumour cells. The second tumour showed distinct papillae covered by layers of polymorphous cells with atypical, often hyperchromatic nuclei. Numerous cells displayed foamy, eosinophilic or clear, sometimes vacuolated cytoplasm. The third case consisted of solid cellular areas composed of pseudostratified columnar cells, most often arranged in perivascular pseudorosette formations. The cells lining papillary structures exhibited marked polymorphism with atypical, often plump nuclei. Mitotic figures were rare and areas of necrosis were observed only in one case. Immunohistochemical staining showed diffuse immunoreactivity for neuron-specific enolase, S-100 protein, cyto-keratin and vimentin. Focal reaction for synaptophysin and chromogranin A and epithelial membrane antigen (EMA) were observed. The tumours lacked expression of GFAP. The Ki-67 labelling index was relatively low but its focal increase was noted in two cases. The final diagnosis of PTPR was based on both predominant papillary morphology and immunohistochemical results. PTPR should be considered in diagnosis of pineal tumours but their natural history, therapeutic strategy and prognosis remain controversial.