Simultaneous extraction and preparation for high-performance liquid chromatography of prostaglandins and phospholipids

Saunders, Royal D.; Horrocks, Lloyd A.

doi:10.1016/0003-2697(84)90559-1

Cited by 87 publications

(39 citation statements)

References 9 publications

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“…The rapid removal and extraction of the tissues prevented postmortem increases in all assayed eico sanoids, as shown by the low levels found in control cer ebra. The extracts were centrifuged to pellet-precipitated protein, passed through 0.65 fLm pore nylon filters, and evaporated under N2• Eicosanoids were separated from other lipids by silicic acid column chromatography (Saunders and Horrocks, 1984), with the eicosanoids being concentrated in the methyl formate fraction. These fractions were evaporated under N2, reconstituted in ra dioimmunoassay buffer, and sampled for quantitation.…”

Section: Methodsmentioning

confidence: 99%

Effects of Cerebral Ischemia and Reperfusion on Prostanoid Accumulation in Unanesthetized and Pentobarbital-Treated Gerbils

Dorman

1988

J Cereb Blood Flow Metab

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Summary: Cerebral ischemia was induced in unanesthe tized gerbils using bilateral carotid artery ligations. The effects of 20 min of global ischemia on the concentrations of prostaglandin F2a (PGF2a), PGE2, 6-keto-prostaglandin Fla (6-keto-PGFla), and thromboxane B2 were deter mined after 0-24 h of reperfusion. Ischemia had little ef fect on eicosanoid production, but significant increases were observed by 5 min of reperfusion, with maximal levels reached by 15 min of reperfusion. PGF2a was the most concentrated prostaglandin in postischemic brain, whereas PGE2 was most concentrated in control cerebra.

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Section: Methodsmentioning

confidence: 99%

Effects of Cerebral Ischemia and Reperfusion on Prostanoid Accumulation in Unanesthetized and Pentobarbital-Treated Gerbils

Dorman

1988

J Cereb Blood Flow Metab

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“…For prostaglandin, phospholipid, and cholesterol analyses, total lipids were extracted with hexane/2-propanol, 3:2 (vol/vol) (28,29), and 20-mg samples were separated on 1.0-g Unisil silicic acid columns. Lipids were eluted with 100 ml each of the following solvents: chloroform (neutral lipids), methyl formate (prostaglandins and cerebrosides), acetone (remaining glycolipids), and methanol (phospholipids).…”

Section: Methodsmentioning

confidence: 99%

Membrane lipid changes in laminectomized and traumatized cat spinal cord.

Demediuk¹,

Saunders²,

Anderson³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

157

105

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Free fatty acid (FFA), diacyiglycerol (acyl2-Gro), icosanoid, phospholipid, and cholesterol levels were measured in samples of cat spinal cord (L2) that were frozen in situ (i) with vertebrae intact, (ii) at various times after laminectomy, and (iu) at various times after laminectomy with compression trauma to the spinal cord. Tissue samples either were grossly dissected into gray and white portions prior to FFA and acyl2Gro analysis or were used whole for the other lipid types. Gray matter total FFA and acyl2Gro values were abnormally high in samples frozen with vertebrae intact and in those frozen 10 min after laminectomy. This indicates that the surgical procedures resulted in some perturbation of spinal cord lipid metabolism. If the experimental animals were allowed to recover for 90 min after laminectomy, the gray matter FFA and acyl2Gro levels were greatly reduced. Compression of the spinal cord with a 170-g weight for 1, 3, or 5mm (following 90 min of recovery after laminectomy) caused significant elevations of total FFA, acyl2Gro, icosanoids, and phosphatidic acid and significant decreases in ethalamine plasmalogens and cholesterol. Among the total FFA, arachidonic acid was found to have the largest relative increase. Comparisons of gray and white matter demonstrate that, in general, changes in white matter FFA and acyl2Gro were similar to those seen in gray matter. However, the increases in white matter levels of FFA and acyl2Gro were delayed, occurring after the elevations in gray matter. For some FFA (e.g., arachidonate), the rise in white matter occurred as gray matter levels were decreasing. This suggests that the initial alteration in spinal cord lipid metabolism after trauma was in gray matter but, with time, spread radially into white matter.

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“…Lipid analysis was performed as previ ously described (Faden et a!., 1990). Briefly, lipids were extracted from brain tissue specimens by homogenizing 5-10 mg samples in 2 ml of hexane/2-propanol (3:2, vol/ vol) (Hara and Radin, 1978;Saunders and Horrocks, 1984). The homogenate was filtered and washed twice with 2 ml of the extracting solvent.…”

Section: Neurochemical Studiesmentioning

confidence: 99%

Effects of Hyperglycemia on the Time Course of Changes in Energy Metabolism and pH during Global Cerebral Ischemia and Reperfusion in Rats: Correlation of¹H and³¹P NMR Spectroscopy with Fatty Acid and Excitatory Amino Acid Levels

Widmer

Abiko

Faden

et al. 1992

J Cereb Blood Flow Metab

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Summary:The effects of hyperglycemia on the time course of changes in cerebral energy metabolite concen trations and intracellular pH were measured by nuclear magnetic resonance (NMR) spectroscopy in rats sub jected to temporary complete brain ischemia. Interleaved 31p and IH NMR spectra were obtained every 5 min be fore, during, and for 2 h after a 30-min bilateral carotid occlusion preceded by permanent occlusion of the basilar artery. The findings were compared with free fatty acid and excitatory amino acid levels as well as with cations and water content in funnel-frozen brain specimens. One hour before occlusion, nine rats received 50% glucose (12 mJJkg i.p.) and five received 7% saline (12 mllkg i.p.). Before ischemia, there were no differences in cerebral metabolite levels or pH between hyperglycemic rats and controls. During the carotid occlusion, the lactatelN acetylaspartate (Lac/NAA) peak ratio was higher (0.73- 456phate were totally depleted in both groups. Within 5-15 min after the onset of reperfusion, the LaclNAA peak ratio increased further in all rats; however, only in ex tremely hyperglycemic rats (serum glucose> 960 mg/dl) did the lactic acidosis progress rather than recover later during reperfusion. Total free fatty acid and excitatory amino acid levels, but not cation concentration or water content, in brain correlated with serum glucose levels during and after ischemia and with NMR findings after 2 h of reperfusion. Although profound hyperglycemia (se rum glucose of 970-1 ,650 mg/dl) appears to be associated with progression of anaerobic glycolysis and failure of cerebral energy metabolism to recover after temporary complete brain ischemia and with postischemic excito toxic and lipolytic reactions thought to participate in de layed cellular injury, severe hyperglycemia (490-720 mg/ dl) was associated with recovery of energy metabolism.

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Simultaneous extraction and preparation for high-performance liquid chromatography of prostaglandins and phospholipids

Cited by 87 publications

References 9 publications

Effects of Cerebral Ischemia and Reperfusion on Prostanoid Accumulation in Unanesthetized and Pentobarbital-Treated Gerbils

Effects of Cerebral Ischemia and Reperfusion on Prostanoid Accumulation in Unanesthetized and Pentobarbital-Treated Gerbils

Membrane lipid changes in laminectomized and traumatized cat spinal cord.

Effects of Hyperglycemia on the Time Course of Changes in Energy Metabolism and pH during Global Cerebral Ischemia and Reperfusion in Rats: Correlation of¹H and³¹P NMR Spectroscopy with Fatty Acid and Excitatory Amino Acid Levels

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Simultaneous extraction and preparation for high-performance liquid chromatography of prostaglandins and phospholipids

Cited by 87 publications

References 9 publications

Effects of Cerebral Ischemia and Reperfusion on Prostanoid Accumulation in Unanesthetized and Pentobarbital-Treated Gerbils

Effects of Cerebral Ischemia and Reperfusion on Prostanoid Accumulation in Unanesthetized and Pentobarbital-Treated Gerbils

Membrane lipid changes in laminectomized and traumatized cat spinal cord.

Effects of Hyperglycemia on the Time Course of Changes in Energy Metabolism and pH during Global Cerebral Ischemia and Reperfusion in Rats: Correlation of1H and31P NMR Spectroscopy with Fatty Acid and Excitatory Amino Acid Levels

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Effects of Hyperglycemia on the Time Course of Changes in Energy Metabolism and pH during Global Cerebral Ischemia and Reperfusion in Rats: Correlation of¹H and³¹P NMR Spectroscopy with Fatty Acid and Excitatory Amino Acid Levels