Methylation subgroup and molecular heterogeneity is a hallmark of glioblastoma: implications for biopsy targeting, classification and therapy.

BACKGROUND: Intratumoral heterogeneity at the cellular and molecular level is a hallmark of glioblastoma (GB) that contributes to treatment resistance and poor clinical outcome. Little is known regarding epigenetic heterogeneity and intratumoral phylogeny and their implication for molecular classification and targeted therapies. PATIENTS AND METHODS: Multiple tissue biopsies (238 in total) were sampled from 56 newly-diagnosed, treatment-naive GB patients from a prospective in-house cohort and publicly available data and profiled for DNA methylation using the Illumina MethylationEPIC array. Methylation-based classification using the glioma classifier developed by Ceccarelli et al. and estimation of the MGMT promoter methylation status via the MGMT-STP27 model were carried out. In addition, copy number variations (CNVs) and phylogeny were analyzed. RESULTS: Almost half of the patients (22/56, 39%) harbored tumors composed of heterogeneous methylation subtypes. We found two predominant subtype combinations: classic-/mesenchymal-like, and mesenchymal-/pilocytic astrocytoma-like. Nine patients (16%) had tumors composed of subvolumes with and without MGMT promoter methylation, whereas 20 patients (36%) were homogeneously methylated, and 27 patients (48%) were homogeneously unmethylated. CNV analysis revealed high variations in many genes, including CDKN2A/B, EGFR, and PTEN. Phylogenetic analysis correspondingly showed a general pattern of CDKN2A/B loss and gain of EGFR, PDGFRA, and CDK4 during early stages of tumor development. CONCLUSIONS: (Epi)genetic intratumoral heterogeneity is a hallmark of GB, both at DNA methylation and CNV level. This intratumoral heterogeneity is of utmost importance for molecular classification as well as for defining therapeutic targets in this disease, as single biopsies might underestimate the true molecular diversity in a tumor.

[Prevention of bicycle accidents]. / Prävention von Fahrradfahrerunfällen.

For a very precise analysis of all injured bicyclists in Germany it would be important to have definitions for "severely injured", "seriously injured" and "critically injured". By this, e.g., two-thirds of surgically treated bicyclists who are not registered by the police could become available for a general analysis. Elderly bicyclists (> 60 years) are a minority (10 %) but represent a majority (50 %) of all fatalities. They profit most by wearing a helmet and would be less injured by using special bicycle bags, switching on their hearing aids and following all traffic rules. E-bikes are used more and more (145 % more in 2012 vs. 2011) with 600,000 at the end of 2011 and are increasingly involved in accidents but still have a lack of legislation. So even for pedelecs 45 with 500 W and a possible speed of 45 km/h there is still no legislative demand for the use of a protecting helmet. 96 % of all injured cyclists in Germany had more than 0.5 ‰ alcohol in their blood, 86 % more than 1.1 ‰ and 59 % more than 1.7 ‰. Fatalities are seen in 24.2 % of cases without any collision partner. Therefore the ADFC calls for a limit of 1.1 ‰. Some virtual studies conclude that integrated sensors in bicycle helmets which would interact with sensors in cars could prevent collisions or reduce the severity of injury by stopping the cars automatically. Integrated sensors in cars with opening angles of 180° enable about 93 % of all bicyclists to be detected leading to a high rate of injury avoidance and/or mitigation. Hanging lamps reduce with 35 % significantly bicycle accidents for children, traffic education for children and special trainings for elderly bicyclists are also recommended as prevention tools. As long as helmet use for bicyclists in Germany rates only 9 % on average and legislative orders for using a helmet will not be in force in the near future, coming up campaigns seem to be necessary to be promoted by the Deutscher Verkehrssicherheitsrat as, e.g., "Helmets are cool". Also, spots in TV should be broadcasted like "The 7th sense" or "Traffic compass", which were warning car drivers many years ago of moments of danger but now they could be used to warn bicyclists of life-threatening situations in traffic.

[Detection of autoantibodies to extractable nuclear antigens in autoimmune diseases of rheumatic origin using immunoblotting--comparison with counter-electrophoresis]. / Nachweis von Autoantikörpern gegen extrahierbare nukleäre Antigene (ENA) bei Autoimmunerkrankungen des rheumatischen Formenkreises mittels Immunoblotting--Vergleich mit der Gegenstromelektrophorese.

Sera from 82 patients with rheumatic autoimmune disease were tested for anti-ENA antibodies by immunoblotting and counterimmunoelectrophoresis (CIE), using HeLa cell extract and rabbit thymus extract, respectively, as antigens. Anti-ENA antibodies were more frequently detected and could be better differentiated by immunoblotting rather than by CIE. There was especially an increase in anti-Sm antibodies (23, in contrast to four positive results), which was only detectable in serum samples for the 59 SLE patients. The sera of the six patients with MCTD all reacted with the 68 U1-RNP antigen. The sera of the five patients with Sjögren's syndrome only recognized the 50k La antigen, while other anti-ENA antibodies were not observed. In SLE anti-La antibodies were often associated with other anti-ENA antibodies. Seven out of eight SLE patients showing a combined detection of antibodies against Sm, U1-RNP and La by immunoblotting demonstrated severe organ involvements, especially lupus nephritis. Therefore, the characterization of anti-ENA antibodies by immunoblotting may contribute to improve the differentiation of connective tissue diseases.