Hypermethylation of regulatory sequences at the locus of the a%-el glutathione S-trnsferase gene GSTPI was detected in 20 of 20 human prostatic carcinoma tissue specimens studied but not in normal tissues or prostatic tissues exhibiting benign hyperplasia. In addition, a striking decrease in GSTPI expression was found to accompany human prostatic MATERIALS AND METHODS Immunohistochemical Staiing for GSTP1. Formalin-fixed paraffin-embedded prostatic tissue sections were stained with anti-GSTP1 antiserum (1:100 dilution; Oncor) using an immunoperoxidase method (Vectastain ABC Kit; Vector Laboratories), with either 3,3'-diaminobenzidine (DAB) or 3-amino-9-ethylcarbazole (AEC) as the peroxidase substrate. As a control for the specificity of staining, tissue sections prepared from normal human liver and normal human kidney were also stained with the anti-GSTP1 antiserum. Each ofthe tissues exhibited the characteristic staining pattern previously described (14).Immunoblot Analysis for GSTP1 Polypeptides. Salinewashed cultured prostatic carcinoma cells (15-19) were collected by centrifugation at 14,000 x g for 10 min at room temperature and then lysed in a protein extraction buffer [2% SDS/101% (vol/vol) glycerol/10 mM dithiothreitol in 62 mM Tris'HCl, pH 7.8] by heating to 950C for 10 min. The DNA content of each cell extract was estimated by using a diphenylamine assay (20). Equivalent extracts from each of the cultured cell lines were subjected to immunoblot analysis for GSTP1 and DNA topoisomerase I polypeptides, using specific antisera (rabbit anti-GSTP1 from Oncor; human antitopoisomerase I provided by W. C. Earnshaw, Johns Hopkins University) as previously described (21).Northern Blot Analyss for GSTPI mRNA. Total RNAs were extracted from cultured cells by the method of Chomczynski and Sacchi (22) and then quantitated by using an orcinol assay (23). Purified RNAs (20 ug) were electrophoresed on 1.5% agarose gels in the presence of2.2 M formaldehyde, transferred to Zeta-Probe (Bio-Rad) filters, and then assessed for GSTPI and TOP] mRNA levels by hybridization with specific 32P-labeled cDNA probes (prepared using the Random Primers DNA Labeling System, GIBCO/BRL). Following hybridization at 50CC for 16 hr (in 50%o formamide/7% SDS/0.5% nonfat dry milk/heat-denatured salmon sperm DNA at 200 pg/ml/300 mM NaCl/2 mM EDTA/20 mM sodium phosphate, pH 7.4), blots were washed with 0.3x SSC (lx SSC is 150mM NaCl/15 mM sodium citrate, pH 7.0) and 0.3% SDS for 60 mi at 650C before being exposed to X-Omat AR (Kodak) film at -700C. Plasmid pGSTii1 containing the entire GSTPI coding sequence (24) was obtained from the American Type Culture Collection; plasmid phTOPl-D1 containing 704 bp of human TOP) cDNA (25) was provided by

Background-Retinal detachment (RD) with proliferative vitreoretinopathy (PVR) often requires surgery. During surgery, a tamponade agent is needed to reduce the rate of recurrent retinal detachment. Objectives-The objective of this review was to evaluate the benefits and adverse outcomes of surgery with various tamponade agents. Search methods-We searched the Cochrane Controlled Register (CENTRAL), MEDLINE, EMBASE, Latin America and Carribbean Health Sciences (LILACS) and the UK Clinical Trials Gateway (UKCTG). There were no language or date restrictions in the search for trials. The electronic databases were last searched on 9 July 2009. Selection criteria-We included randomized clinical trials comparing patients treated with various tamponade agents.

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