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The assay was conducted in the wells of 96-well black plastic microplates, in which 20 L of enzyme solution and 60 L of bile salt were mixed together, and then 20 L of substrate was added, with mixing, at the start of the reaction, which proceeded at room temperature for up to 30 min, with measurement at various intermediate time points

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By addressing the underlying causes rather than merely alleviating symptoms, CBT-I is a sustainable and highly effective treatment option for insomnia

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