Erlosamide or unacceptable toxicity occurred; additional treatment beyond 12 months was allowed at the discretion of the treating physician, assuming no significant toxicity, and with the consent of the patient. Patient Evaluations Pretreatment evaluation included a complete history and physical and neurologic examination. Prestudy laboratory tests, obtained within 14 days after treatment, included a complete blood count with differential and serum creatinine, total bilirubin, aspartate transaminase, alanine aminotransferase, alkaline phosphatase, blood urea nitrogen, glucose, potassium, sodium, and anticonvulsant levels ; serum pregnancy tests were performed for women of childbearing potential. Pathology slides from the most recent surgical material were submitted for retrospective pathology review to confirm the diagnosis and to evaluate molecular abnormalities in the tumor.
During RT, a CBC and Ramelteon clinical trial differential were performed every 2 weeks, then at weeks 3 and 4 after raltegravir structure the start of each 28 day temozolomide cycle. Blood chemistry tests were performed every 4 weeks. Safety evaluations were performed using the Common Terminology Criteria for Adverse Events . Brain MRI was performed at baseline , 2 3 weeks after the completion of RT, and then every 8 weeks while patients were receiving treatment. The Macdonald Criteria were used to evaluate radiologic progression.20 All patients were observed for OS. Patients who experienced disease progression were observed for survival every 3 months.
Pharmacogenomics Genomic DNA was isolated from formalin fixed, paraffin embedded tumor tissue or from blood samples with use of standard techniques Aprepitant solubility and was subjected to bisulfite treatment, as described elsewhere.21,22 TheO 6 methylguanine DNAmethyltransferase methylation specific polymerase chain reactions were performed using a 2 step approach, and the results were confirmed by a 1 step MSP in a subset of tumors.23 26 Fetal brain tissue was used to generate positive and negative controls for the MSP with native fetal brain DNA positive for the unmethylated polymerase chain reaction product and SssI treated in vitro methylated fetal brain DNA serving as a positive control for the methylated MGMT PCR product. The PCR products were resolved on 4% agarose gels. Analysis of MSP data was performed by investigators who were blinded to the clinical data.
Immunohistochemistry was used to evaluate molecular markers, including epidermal growth factor receptor , phosphatase and tensin homolog , phosphorylated S6 ribosomal protein using the 2211 and 2215 antibodies, glycogen synthase kinase 3 affirmative team beta , phosphorylated cAMP response element binding protein , VEGF, and mitogen activated protein kinase . The IHC assays were scored using a 0 to +3 scoring system. No positive staining was scored 0; at least 25% immunoreactivity of cells was scored +1; 26% 75% was scored +2; and ≥76% was scored +3. Results were analyzed using the actual IHC score, and the level of positivity was included as part of the assessment. Thus, any level of positivity was considered to be positive, but the range was taken into account. Statistical Plan The initial calculation of sample size was based on the goal of increasing survival compared with the historical median survival time of 15 months.