Dynamic expression of 11 miRNAs in 83 consecutive primary and corresponding recurrent glioblastoma: correlation to treatment, time to recurrence, overall survival and MGMT methylation status.

Background Glioblastoma (GBM) is the most common and the most malignant glioma subtype. Among numerous genetic alterations, miRNAs contribute to pathogenesis of GBM and it is suggested that also to GBM recurrence and resistance to therapy. Based on publications, we have selected 11 miRNAs and analyzed their expression in GBM. We hypothesized that selected miRNAs are differentially expressed and involved in primary as well as in recurrent GBM, that show significant expressional differences when different treatment options are in question, and that are related to certain patients and tumor characteristics. Patients and methods Paraffin embedded tissues, obtained from primary and corresponding recurrent tumor from 83 patients with primary GBM were used. Eleven miRNAs (miR-7, miR-9, miR-15b, miR-21, miR-26b, miR-124a, miR-199a, let-7a, let-7b, let-7d, and let-7f) were selected for qPCR expression analysis. For patients who received temozolamide (TMZ) as chemotherapeutic drug, O6-methylguanine-DNA methyltransferase (MGMT) methylation status was defined using the methyl-specific PCR. Results There was a significant change in expression of miR-7, miR-9, miR-21, miR-26b, mirR-124a, miR-199a and let-7f in recurrent tumor compared to the primary. In recurrent tumor, miR-15b, let-7d and let-7f significantly changed comparing both treatment options. We also observed difference in progression free survival between patients that received radiotherapy and patients that received radiotherapy and chemotherapy, and longer survival for patients who received chemotherapy after second surgery compared to not treated patients. miR-26b showed correlation to progression free survival and let-7f to overall survival. We did not find any expression difference between the tumors with and without methylated MGMT. Conclusions Our data suggest that analyzed miRNAs may not only contribute to pathogenesis of primary GBM, but also to tumor progression and its recurrence. Moreover, expression of certain miRNAs appears to be therapy-dependent and as such they might serve as additional biomarker for recurrence prediction and potentially predict a therapy-resistance.

Identifying Novel Glioma-Associated Noncoding RNAs by Their Expression Profiles.

Long noncoding RNAs (lncRNAs) and microRNAs (miRNAs) play a significant role in cancer development as regulators of protein-coding genes. Their dysregulation was in some extent already associated with glioma, the most aggressive primary brain tumours in adults. The correct diagnosis and treatment selection due to high tumour heterogeneity might be difficult and inadequate, resulting in poor prognosis. Studies of expression patterns of noncoding RNAs (ncRNAs) could provide useful insight in glioma molecular development. We used the qPCR approach to screen and investigate the expression of lncRNAs that were previously deregulated in other cancer types. The study showed altered expression levels for numerous lncRNAs across histologically different glioma samples. Validation of few lncRNAs showed association of expression levels with histological subtype and/or malignancy grade. We also observed deregulated and subtype-distinctive expression for four lncRNA-associated miRNAs. Expression of few lncRNAs and miRNA was also associated with patients' survival, showing potential prognostic value. Several ncRNAs, some already related to glioma and some, to the best of our knowledge, investigated for the first time, might be of greater importance in glioma molecular development and progression. Finding the subtype-specific lncRNA and/or miRNA expression patterns may contribute additional information for a more objective classification.

Expression of LOC285758, a Potential Long Non-coding Biomarker, is Methylation-dependent and Correlates with Glioma Malignancy Grade.

BACKGROUND: Identifying the early genetic drivers can help diagnose glioma tumours in their early stages, before becoming malignant. However, there is emerging evidence that disturbance of epigenetic mechanisms also contributes to cell's malignant transformation and cancer progression. Long non-coding RNAs are one of key epigenetic modulators of signalling pathways, since gene expression regulation is one of their canonical mechanisms. The aim of our study was to search new gliomagenesis-specific candidate lncRNAs involved in epigenetic regulation. PATIENTS AND METHODS: We used a microarray approach to detect expression profiles of epigenetically involved lncRNAs on a set of 12 glioma samples, and selected LOC285758 for further qPCR expression validation on 157 glioma samples of different subtypes. To establish if change in expression is a consequence of epigenetic alterations we determined methylation status of lncRNA's promoter using MS-HRM. Additionally, we used the MLPA analysis for determining the status of known glioma biomarkers and used them for association analyses. RESULTS: In all glioma subtypes levels of LOC285758 were significantly higher in comparison to normal brain reference RNA, and expression was inversely associated with promoter methylation. Expression substantially differs between astrocytoma and oligodendroglioma, and is elevated in higher WHO grades, which also showed loss of methylation. CONCLUSIONS: Our study revealed that lncRNA LOC285758 changed expression in glioma is methylation-dependent and methylation correlates with WHO malignancy grade. Methylation is also distinctive between astrocytoma I-III and other glioma subtypes and may thus serve as an additional biomarker in glioma diagnosis.

Differentially expressed proteins in glioblastoma multiforme identified with a nanobody-based anti-proteome approach and confirmed by OncoFinder as possible tumor-class predictive biomarker candidates.

Glioblastoma multiforme is the most frequent primary malignancy of the central nervous system. Despite remarkable progress towards an understanding of tumor biology, there is no efficient treatment and patient outcome remains poor. Here, we present a unique anti-proteomic approach for selection of nanobodies specific for overexpressed glioblastoma proteins. A phage-displayed nanobody library was enriched in protein extracts from NCH644 and NCH421K glioblastoma cell lines. Differential ELISA screenings revealed seven nanobodies that target the following antigens: the ACTB/NUCL complex, VIM, NAP1L1, TUFM, DPYSL2, CRMP1, and ALYREF. Western blots showed highest protein up-regulation for ALYREF, CRMP1, and VIM. Moreover, bioinformatic analysis with the OncoFinder software against the complete "Cancer Genome Atlas" brain tumor gene expression dataset suggests the involvement of different proteins in the WNT and ATM pathways, and in Aurora B, Sem3A, and E-cadherin signaling. We demonstrate the potential use of NAP1L1, NUCL, CRMP1, ACTB, and VIM for differentiation between glioblastoma and lower grade gliomas, with DPYSL2 as a promising "glioma versus reference" biomarker. A small scale validation study confirmed significant changes in mRNA expression levels of VIM, DPYSL2, ACTB and TRIM28. This work helps to fill the information gap in this field by defining novel differences in biochemical profiles between gliomas and reference samples. Thus, selected genes can be used to distinguish glioblastoma from lower grade gliomas, and from reference samples. These findings should be valuable for glioblastoma patients once they are validated on a larger sample size.

Neurosurgical training programme in selected European countries: from the young neurosurgeons' point of view.

AIM: In this paper we discuss the Neurosurgical Training Programme (NTP) in some European countries. MATERIAL AND METHODS: Although there is no official data on how many neurosurgeons are certified in Europe, our calculation shows that this number is somewhat lower than in the United States of America and even 3 times lower than in Japan (per 100.000 population). It is also evident that there is no consensus in the Programme duration or in the official NTP content, despite the recommendations of the EANS (The European Association of Neurosurgical Societies). Trainees from outside the European Union (EU) are under-represented in the EANS training courses. We believe that in the eastern part of Europe there is the most space for improvement in neurosurgical training. Solving of all these problems requires first and foremost their recognition and consideration - then devising a solution. CONCLUSION: The purpose of this paper is to compare and contrast several NTP's in Europe in order to promote a more coherent medical education. Some remarks and suggestions from the perspective of young neurosurgeons are given.

Use of 3D visualisation of medical images for planning and intraoperative localisation of superficial brain tumours: our experience.

BACKGROUND: In the present article, we assessed the role, adequacy and application accuracy of intraoperative visual guidance based on the computer 3D visualisation of preoperative medical images in the surgery of superficial brain tumours. MATERIALS AND METHODS: For 30 consecutive patients with convexity meningioma or cortical/subcortical brain tumour, we used 3D visualisation of post-contrast fast spoiled gradient recalled (FSPGR) MR images to plan optimal positions for the trepanation opening and/or corticotomy site. At the beginning of the surgery, planned positions were transferred to the scalp and the cortical surface of the patient by visually matching the 3D surfaces with the operative field. The feasibility of visual matching was assessed by counting the number of cases in which this was possible. On the exposed cortical surface, we measured the mismatch between the centre of the actual trepanation opening and the planned corticotomy site, where possible. RESULTS: During computer-assisted 3D planning, the centre of the trepanation opening, initially defined on the basis of 2D diagnostic images, was redefined in all our cases by an average repositioning distance of 19.7 mm +/- 7.6 mm. During surgery, the transfer of the planned centre of the trepanation opening and the corticotomy site was possible in all (30/30) and in 70% (19/27) of the cases, respectively. Where assessable, the mismatch between the centre of the actual trepanation opening and the planned corticotomy site was less than 1 cm in 70% of cases (12/17) and more than 2 cm in 6% (1/17) of cases. CONCLUSIONS: Intraoperative visual guidance based on 3D visualisation proved to be adequate and accurate for locating superficial brain tumours in cases where transfer of planned surgical targets to the surgical field was possible. Decision about its use should be based on preoperative computer-assisted 3D planning, in which the feasibility of visual matching during surgery can and must be assessed.

Prevention of nosocomial lower respiratory tract infections in patients after intracranial artery aneurysm surgery with a short course of antimicrobials.

Nosocomial infections at a department of neurosurgery were followed prospectively. In 1999, a high incidence of nosocomial lower respiratory tract infections (NLRTI) was observed in patients after intracranial artery aneurysm surgery (ICAAS). From February to December 2000, a short course of ciprofloxacin and co-trimoxazole was given prophylactically to all patients with ICAAS. The incidence of nosocomial infections in patients after ICAAS fell from 78.4 to 30.9% (P<0.0001). The incidence of NLRTI fell from 43.3 to 13.6% (P<0.0001) and the incidence of urinary tract infections from 12.4 to 2.5% (P=0.015). No significant change in antibiotic sensitivity at the department was observed. Antibiotic use decreased from 100.4 defined daily doses (DDD) to 85.4 DDD per 100 bed-days.