5 C s 1, with the signal acquisition mode set to continuous. Quantification of data was performed using the computer program LinReg PCR in which linear re gression on the Log per cycle number data is applied to determine the amplification efficiency per sample. truly The starting concentration of each specific product was divided by the starting concentration of reference genes and this ratio was compared between patientcontrol groups. Human material The human cases included in this study were obtained from the files of the departments of neuropathology of the Academic Medical Center and the VU University Medical Center, both situated in Amsterdam and both ter tiary referral centers for brain tumor patients in the Netherlands. We examined immunocytochemically Inhibitors,Modulators,Libraries 73 surgical specimens of brain tumor patients with astro cytic tumors.
Normal appearing control cortex white matter was obtained at autopsy from eight adult control patients without a history Inhibitors,Modulators,Libraries of seizures or other neurological diseases. All autopsies were performed within 12 h after death. Cortical samples of five patients with non glial brain tumors and without refractory epilepsy were also analyzed. Frozen tis sue from histologically normal cortex and GBM samples was used for western blot analysis and total RNA prepared from normal cortex and GBM was used for qPCR. A chart review was conducted of all patients. Epilepsy was defined as the experience of one or more seizures and data regarding seizure frequency and seizure type were obtained from patient histories. We collected additional data including age, gender, tumor location, and Inhibitors,Modulators,Libraries epilepsy duration.
Informed consent was obtained for the use of brain tissue and for access to medical records for research purposes. Tissue Inhibitors,Modulators,Libraries was obtained and used in a manner compliant with the Declaration of Hel sinki. Two neuropathologists reviewed all cases inde pendently and the diagnosis was confirmed according to the revised WHO classification of tumors of the central nervous system. Tissue preparation for immunocytochemistry Tissue was fixed in 10% buffered formalin and embed ded in paraffin. Paraffin embedded tissue was sectioned at 5 um, mounted on precoated glass slides and used for immunohistochemical staining as described below. Antibodies Antibodies specific for glial fibrillary acidic protein, vimentin, neuronal nuclear protein, synaptophysin, Ki67, DP, DQ, DR, MAP2 and p53, were used in the routine immunohistochemical analysis of glial tumors.
For the detection of Kir4. 1, we used a polyclonal rabbit Inhibitors,Modulators,Libraries antibody for the detection of IL 1B a polyclonal goat anti body and for the detection of HMGB, we used a polyclonal rabbit antibody. Immunohistochemistry Paraffin embedded sections were deparaffinized, re hydrated, and incubated for 20 min in 0. 3% H2O2 diluted in methanol to quench the download catalog endogenous per oxidase activity.