A fast, nondestructive purification scheme for prostaglandin H2 using a nonaqueous, bonded-phase high-performance liquid chromatography system

Zulak, I. M.; Puttemans, M.L.; Schilling, Alexander; Hall, Elizabeth R.; Venton, D. L.

doi:10.1016/0003-2697(86)90509-9

Cited by 8 publications

(4 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Human lymphocytes are capable of taking up arachidonic acid (AA)' and incorporating it into their phospholipids, yet they appear to possess minimal ability to convert that arachidonate into biologically active metabolites (1,2 Receivedfor publication 5 May 1986 and in revisedform 13 January 1987. 1.…”

Section: Introductionmentioning

confidence: 99%

“…PGH2 purification. PGH2 was prepared using AA and ram seminal vesicle microsomes, and the biosynthetically produced PGH2 was purified over a gravity flow silicic acid column and a cyanobonded phase HPLC self-packed column using a hexane-isopropanol gradient (5). The purified product is free from other eicosanoids as analyzed by gas chromatography-mass spectrometry.…”

Section: Introductionmentioning

confidence: 99%

“…To obtain washed buffy coat preparations, venous blood was drawn into tubes containing 1/10 vol of 3.8% sodium citrate, mixed and centrifuged at 3,000 g for 20 min. After removing plasma, the buffy-coat layer was carefully separated and washed three times in Tyrode's buffer and the final preparation was suspended in 2 ml Tyrode's buffer that contained 5 (8). Recovery for eicosanoids was -95%.…”

Section: Introductionmentioning

confidence: 99%

“…These results suggest that lymphocytes do not metabolize AA directly, but that they possess sufficient PGI2 synthase activity to synthesize PG12 from PGH2. To test this hypothesis, 5 ,uM ultrapurified PGH2 was incubated with washed lymphocytes. The supernatant was applied to HPLC, and 6KPGFIa content in the HPLC fractions was measured by RIA.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Interaction between lymphocytes and platelets in the synthesis of prostacyclin.

Wu¹,

Papp²,

Manner³

et al. 1987

J. Clin. Invest.

View full text Add to dashboard Cite

To test the hypothesis that prostacyclin (PGI2) is formed via a biochemical interaction between platelets and lymphocytes, we measured eicosanoids by high performance liquid chromatography (HPLC) and radioimmunoassay (RIA). A distinct 6-keto-prostaglandin Fl, (6KPGF1,,) peak was noted when [14Clarachidonic acid ([14CjAA) was added to the mixed cell preparations which was increased by pretreating platelets with 1-benzylimidazole (1-BI). Lymphocytes prelabeled with [14CIAA failed to form 6KPGF1, when stimulated with phytohemagglutinin (PHA) or ionophore A23187. When the prelabeled platelets were suspended together with aspirin-treated lymphocytes and stimulated with ionophore, thrombin, or collagen, a 6KPGF1, peak was detected and enhanced by 1-BI. These results were supported by quantifying the 6KPGF1, content in the HPLCpurified fraction by RIA. Adding prostaglandin H2 (PGH2) directly to lymphocytes led to 6KPGF1,, production. Platelet aggregation and release were inhibited by lymphocytes in a doserelated manner. We conclude that lymphocytes possess PG12 synthase activity which is capable of converting platelet-derived PGH2 into PGI2. PGI2 formed is sufficient to inhibit platelet function.

show abstract

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations