This study was done to evaluate various DNA and RNA extractions from archival FFPE tissues. A total of 30 FFPE blocks from the years of 2004 to 2006 were assessed with each modified and adapted method. Extraction protocols evaluated include the modified enzymatic extraction method (Method A), Chelex-100 extraction method (Method B), heat-induced retrieval in alkaline solution extraction method (Methods C and D) and one commercial FFPE DNA Extraction kit (Qiagen, Crawley, UK). For RNA extraction, 2 extraction protocols were evaluated including the enzymatic extraction method (Method 1), and Chelex-100 RNA extraction method (Method 2). Results show that the modified enzymatic extraction method (Method A) is an efficient DNA extraction protocol, while for RNA extraction, the enzymatic method (Method 1) and the Chelex-100 RNA extraction method (Method 2) are equally efficient RNA extraction protocols.

INTRODUCTION: Fungal peritonitis is uncommon but important complication of continuous ambulatory peritoneal dialysis (CAPD). It accounts for 5 percent-15 percent of the total burden of peritonitis in most CAPD units. At the National Kidney and Transplant Institute (NKTI), the incidence increased from 11 percent in 1997 to 21 percent in 2000. Since this complication is associated with severe morbidity, CAPD drop-out and death, measures to decrease or prevent its occurrence are necessary.

OBJECTIVES: (l) To determine if nystatin is effective as prophylaxis for fungal peritonitis in patients with an ongoing CAPD-related bacterial peritonitis; and (2) to determine the predisposing factors for fungal peritonitis

DESIGN: Randomized single-blinded trial

SETTING: National Kidney and Transplant Institute, tertiary renal referral center in the Philippines

METHODS: From June 2001 to December 2004, newly diagnosed patients with end-stage renal disease (ESRD) on CAPD were randomized to receive nystatin as prophylaxis for fungal peritonitis during their first bacterial peritonitis episode and for all successive episodes within the next 6 months. Group 1 patients received nystatin concomitantly with the antibiotics for bacterial peritonitis while Group 2 received antibiotics alone. The incidence and the time to development of fungal peritonitis were measured. Intention-to-treat analysis was used.

RESULTS: There were 133 eligible patients, of whom 74 patients were randomized to Group 1 and 59 patients were randomized to Group 2. Eighty-three patients completed the study: 47 in Group 1 and 36 in Group 2. The cumulative incidence of developing fungal peritonitis was 10.8 percent. Group 1 had a slightly lower incidence of fungal peritonitis compared to Group 2 (10.6 percent vs 11.1 percent) but this was not statistically significant (p-1.0). Nystatin prophylaxis delayed the time to develop fungal peritonitis by 5 months but this was not statistically significant (p-0.94). Among the potential risk factors studied, only the presence of diabetes mellitus was associated with the development of fungal peritonitis.

CONCLUSION: There were no significant differences in clinical outcomes in the two treatment groups. However, there appeared to be a delay in the development of fungal peritonitis with the use of nystatin prophylaxis within the first 5 months of a bacterial peritonitis episode. (Author)

Fungal Peritonitis ; Nystatin ; Peritoneal Dialysis, Continuous Ambulatory ; Peritonitis