Reliable, sensitive, rapid and quantitative enzyme-based assay for gamma-hydroxybutyric acid (GHB)

Several assays for gamma-hydroxybutyrate (4-hydroxybutyrate, GHB) have been developed based on the enzyme gamma-hydroxybutyrate dehydrogenase (GHB-DH). Enzymatic oxidation of GHB by NAD+ is coupled to diaphorase-mediated reduction of pro-dye to yield colored product. GHB-DH from Ralstonia eutropha was cloned and expressed as a stable fusion protein easily purified by affinity chromatography. Quantitative initial velocity and endpoint versions of the assay in solution are described. Michaelis-Menten parameters for oxidation of GHB and ethanol were estimated. A semi-quantitative "dipstick" version of the assay on paper also is described. Both solution endpoint and "dipstick" assays are sensitive to about 0.05 mg GHB/mL using 10 microL of sample. Ethanol at concentrations possible in urine and agents used to stabilize physiological fluids for forensics analysis do not interfere significantly. The "dipstick" assay also allows detection of GHB in alcoholic beverages after evaporation of about one-fourth drop of beverage before testing. The enzymatic assay for GHB is reliable, sensitive, inexpensive and rapid.     

La subsidiarité en droit européen et canadien : Une comparaison

The principle of subsidiarity, one of the most important in European Union (EU) law and entrenched in the EU treaties, has recently entered the constitutional discourse in Canada thanks to some Supreme Court decisions regarding division of powers. After a brief overview of the intellectual history of subsidiarity, this article offers a comparison of how subsidiarity has been dealt with by the Supreme Court of Canada and by the European Court of Justice. The ultimate goal is to see what lessons Canada can draw from the EU's longstanding experience with subsidiarity, particularly in the ambit of social policy.     

Choice of Law in Tort—Blending in with the Landscape of the Conflict of Laws?

The reformed choice of law in tort rules contained in the Private International Law (Miscellaneous Provisions) Act 1995 have proved a source of great controversy. This piece attempts to show that there are wider concerns fuelled by the Act. It seeks to demonstrate that one can not reform one area of the Conflict of Laws (such as choice of law in tort) in isolation from other areas (such as jurisdiction in tortious matters and choice of law in contract) and to illustrate the unsatisfactory state that the statutory reforms have had on these other areas. The analysis of the problems generated by the Act may be used as a model for a far more fundamental proposition: namely that the tendency to regard choice of law and jurisdiction as separate areas throughout the Conflict of Laws, rather than as combining to form a coherent law of international litigation, is a critical shortcoming of the study of the subject.     

Justice, at last, in the archetypical patent entitlement dispute

The Court of Appeal has held that Cinpres was not estopped fromsucceeding in its entitlement claim, despite having failed inan earlier claim concerning the related application, and isthe rightful owner of a disputed European patent.