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For IC 50 determination and kinetic experiments, the enzyme stock (2 mg/mL) was diluted 20-fold or 35-fold with 100 mM sodium phosphate buffer, pH 6.5, containing 1 mM EDTA and 0.05% (w/v) BSA, respectively

Antenatal diagnosis Antenatal diagnosis of 5-oxoprolinase deficiency has not been reported to date

Synthesised genes were sub-cloned into a pET21a (Novagen) vector with a C-terminal 6His tag

The combination of reduced intake and impaired absorption creates a trajectory toward deficiency over months and years of treatment

Nonetheless, optimal doses, treatment schedules and testing efficacy of other antioxidants in preventing THC-induced abnormalities remain to be established