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Two possibilities are that DHA concentrations are extremely low in compartments such as the chloroplast and cytosol or that rate constants for the chemical reaction are much slower in the crowded cellular environment than those measured in dilute solutions in vitro, making enzymes necessary to favour the reaction between DHA and GSH

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Optional: Freezing the cell pellet at -20 C for 30 min prior to incubation at room temperature improves lysis by lysozyme

This mechanism relies on intact pituitary and hypothalamic function, making efficacy dependent on baseline hormonal reserve and individual receptor sensitivity

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