Many methods for the detection of genomic DNA methylation states have appeared. Currently, nearly all such methods employ bisulfite-mediated deamination of denatured DNA. While this treatment effectively deaminates cytosines to uracils, leaving most 5-methylcytosines intact, it also introduces abasic sites that generate a significant number of single-strand breaks in DNA. We have investigated the interplay of these two processes in order to determine their relative effects on the methylation-sensitive QPCR method. The extent of cleavage of the input DNA is significant and appears to be an increasing function of DNA concentration. Even so, the results suggest that only ∼10% of a 62-nt target will be lost due to degradation and targets up to 131 nt will suffer only a 20% loss. More significant losses were found to occur during the subsequent removal of bisulfite and desulfonation steps that appear to be the result of size selectivity associated with matrix binding and elution required prior to QPCR in the most commonly used protocols. For biospecimens yielding <1 μg of DNA, these findings suggest that bisulfite treatment, in current implementations of MS-QPCR, result in low recoveries that preclude reliable analysis of DNA methylation patterns regardless of target size.

Prostate cancer is extremely rare in men under 35 years of age. The tumour is invariably poorly differentiated and aggressive, with rapidly growing bulky soft tissue metastases and negative tumour markers. Bone metastases develop late and are usually osteolytic. The disease responds poorly to radiation or hormonal therapy and is too advanced at presentation for radical surgery. Chemotherapy appears to be of some benefit, though in the majority of cases death occurs within a year. We describe a 31-year-old man with carcinoma of the prostate. A review of the literature is presented. This is the first patient in whom the epithelial origin of the prostate cancer was confirmed by immunoperoxidase staining with prostatic specific antigen. Plasma prostatic specific antigen was normal despite a large tumour burden and widespread metastases. He did not respond to conventional treatment. The phenotypic expression and biological behaviour of these tumours are distinct from those occurring in men beyond the fourth decade.

The Stamey vesical neck suspension has a good early success rate but the results in the longer term are not acceptable. This operation should not be used as a first-line treatment for stress incontinence and should be reserved for specific patient subgroups.

BACKGROUND: TMPRSS2:ERG fusions are promising prostate cancer biomarkers. Because they can occur in multiple forms in a single cancer specimen, we developed a quantitative PCR test that detects both type III and type VI TMPRSS2:ERG fusions. The assay is quantified from a standard curve determined with a plasmid-cloned type III TMPRSS2:ERG fusion target.

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