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Carla S. Jung, Brian A. Iuliano, Judith Harvey-White, Michael G. Espey, Edward H. Oldfield, and Ryszard M. Pluta

vasospasm remained unchanged after SAH. Levels of ADMA in animals with and without vasospasm were significantly different on Day 7 (#p < 0.01) and Day 14 (#p < 0.04). Asymmetric dimethyl- l -arginine levels in serum of the control group (5.8 ± 2.9 µM) were comparable tothose detected in animals in the SAH group on all days after SAH (Days 0–21: mean 7.3 ± 5.8 µM, median 4.8 µM, and range 1.1–24.8 µM; p = 0.43). Levels of ADMA serum in animals with and without vasospasm remained essentially unchanged and did not demonstrate any significant difference on Day 7 (7

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Ryszard M. Pluta, Carla S. Jung, Judith Harvey-White, Anne Whitehead, Sabrina Shilad, Michael G. Espey, and Edward H. Oldfield

(Waters Corp., Milford, MA) was used. Asymmetric Dimethyl l -Arginine Measurements Asymmetric dimethyl l -arginine and SDMA (the inactive enantiomer of ADMA) levels in the media and cell lysate samples, as well as in the serum and CSF samples, were assessed using HPLC. One hundred microliters of media/cell lysate, 80 µl of each serum/CSF sample, and 50 µM methionine sulfone, added as an internal standard to each sample, were derivatized using phenylisothiocyanate and dried under nitrogen. Ten-microliter aliquots of each sample were then analyzed using HPLC. The

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Carla S. Jung, Edward H. Oldfield, Judith Harvey-White, Michael G. Espey, Michael Zimmermann, Volker Seifert, and Ryszard M. Pluta

because no significant reduction in vessel diameter, and therefore in vasospasm, was detectable. Analysis of CSF Asymmetric Dimethyl- l -Arginine, l -Arginine, and l -Citrul-line Levels High-performance liquid chromatography, a widely applied method that has good analytical sensitivity and accuracy, was used to determine levels of ADMA, l -arginine, and l -citrulline in the CSF and serum of all patients, as described previously. 5 , 20 , 27 , 31 , 47 In short, 50 μM methionine sulfone was added to each sample as an internal standard and 80 μl of the CSF

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Ali Reza Fathi, Ryszard M. Pluta, Kamran D. Bakhtian, Meng Qi, and Russell R. Lonser

indices . Neurosurgery 42 : 1076 – 1082 , 1998 10 Hoh BL , Ogilvy CS : Endovascular treatment of cerebral vasospasm: transluminal balloon angioplasty, intra-arterial papaverine, and intra-arterial nicardipine . Neurosurg Clin N Am 16 : 501 – 516 , vi , 2005 11 Jung CS , Iuliano BA , Harvey-White J , Espey MG , Oldfield EH , Pluta RM : Association between cerebrospinal fluid levels of asymmetric dimethyl-L-arginine, an endogenous inhibitor of endothelial nitric oxide synthase, and cerebral vasospasm in a primate model of subarachnoid