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Layer 5 ml of the coelomocyte-containing fluid on top of the gradient and centrifuge at 1500 g for 30 min at 4C (see Note 9)

The prevalence of B12 deficiency increases significantly after age 60, and correcting deficiency can help maintain normal neurological function

Analysis of tryptophan was performed after separation of the alkaline digest on a C18 column (Phenomenex, Torrance, CA, USA) using isocratic elution (1.0 mL/min flow rate at 28 C and total run time of 34 min)

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