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A high-throughput mass spectrometry assay to measure the catalytic activity of phosphatidylserine decarboxylase (PISD) is described. PISD converts phosphatidylserine to phosphatidylethanolamine during lipid synthesis. Traditional methods of measuring PISD activity are low throughput and unsuitable for the high-throughput screening of large compound libraries. The high-throughput mass spectrometry assay directly measures phosphatidylserine and phosphatidylethanolamine using the RapidFiretrade mark platform at a rate of 1 sample every 7.5 s. The assay is robust, with an average Z' value of 0.79 from a screen of 9920 compounds. Of 60 compounds selected for confirmation, 54 are active in dose-response studies. The application of high-throughput mass spectrometry permitted a high-quality screen to be performed for an otherwise intractable target.

Stroke disease remains a national priority due to the high morbidity and mortality. Good clinical practice dictates that risk factors are identified and corrected. Various therapeutic regimens have been tested in trials of sufficient strength to give answers. In the second European Stroke Prevention Study (ESPSZ), low dose aspirin (50 mg/d), sustained release dipyridamole (200 mg/d) were shown to reduce recurrence of stroke and TIA. The combination was twice as effective. Also in the Clopidogrel/aspirin Prevention of Recurrence of Ischaemic Events (CAPRIE) trial clopidogrel was shown to be superior to aspirin. In clinical practice patients with stroke or TIA should be investigated rapidly, their risk factors corrected and appropriate drug regimens implemented.

In a randomized double-blind trial, patients with ischemic rest pain of the leg received 8 days of treatment with intravenous injections of the defibrinating agent, ancrod (Arvin) or saline. Plasma fibrinogen, plasma viscosity, and blood viscosity were significantly reduced during ancrod therapy. After 27 patients had completed treatment, sequential analysis showed no preference for either therapy. In each group two-thirds of the patients claimed improvement in pain after treatment. There were no significant differences between treatment groups in analgesic consumption, Doppler systolic pressure ratio, or surgical intervention in the 6 months following treatment. We conclude that in patients with ischemic rest pain, ancrod therapy is not superior to placebo injections, and that the effect of placebo treatment is considerable.

Nine patients with extensive peripheral arterial disease were treated with subcutaneous injections of ancrod (Arvin) for 10 to 21 days. Reduction in plasma fibrinogen was associated with a sustained reduction in plasma and blood viscosity, and a sustained increase in nutritional skin blood flow, measured by a Xenon-133 clearance technique (P less than 0.001). These findings may be relevant to the therapeutic effect of ancrod in ischemic rest pain.