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T R Pettitt

Publications and source records attributed to T R Pettitt.

23 records · Page 2Linked to original sources

Synthesis of lipoxins and other lipoxygenase products by macrophages from the rainbow trout, Oncorhynchus mykiss.

Rainbow trout macrophages maintained in short term culture when incubated with either calcium ionophore, A23187, or opsonized zymosan synthesize a range of lipoxygenase products including lipoxins and leukotrienes. These cells are unusual in that they generate more lipoxin than leukotriene following such challenge. The main lipoxin synthesized was lipoxin (LX) A4. This compound was identified by cochromatography with authentic standard during reversephase high performance liquid chromatography, by ultra violet spectral analysis, radiolabeling following incorporation of [14C]arachidonic acid substrate into macrophage phospholipids, and gas chromatography electron impact mass spectrometry of the methyl ester, trimethylsilyl ether derivative. Other 4-series lipoxins synthesized by trout macrophages were identified as 11-trans-LXA4, 7-cis-11-trans-LXA4, and 6(S)-LXA4. These cells also produced 5-series lipoxins tentatively identified as LXA5, 11-trans-LXA5 and possibly 6(S)-LXA5. No LXB4 or LXB5 was, however, detected. The dynamics of leukotriene and lipoxin release were also determined. Lipoxin generation was slower than leukotriene generation the latter reaching a maximum after 30 min of exposure to ionophore (5 microM, 18 degrees C) compared with 45 min for the former.

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Fatty acid composition and lipoxygenase metabolism in blood cells of the lesser spotted dogfish, Scyliorhinus canicula.

1. The fatty acid composition of erythrocytes and leucocytes of the elasmobranch, Scyliorhinus canicula, was determined so as to indicate substrate availability for eicosanoid formation. 2. Leucocytes showed a greater degree of fatty acid unsaturation than the erythrocytes, with particularly high levels of docosahexaenoic acid (22:6,n-3). 3. The major eicosanoid precursors, arachidonic acid (20:4,n-6) and eicosapentaenoic acid (20:5,n-3), represented 13.9% and 5.2% of the total fatty acid, respectively, in erythrocytes compared with 10.7% and 6% in leucocytes. 4. Whole blood and isolated leucocytes were stimulated with calcium ionophore, A23187 and the resulting lipoxygenase products separated by reverse phase high performance liquid chromatography. 5. The main lipoxygenase products formed were 6-trans-leukotriene B4, 6-trans-12-epi-leukotriene B4, 5(S),6(R) dihydroxyeicosatetraenoic acid and 5- and 15-hydroxyeicosatetraenoic acid. 6. No leukotriene B4, leukotriene B5, or lipoxins were detected.

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Lipoxins are major lipoxygenase products of rainbow trout macrophages.

Rainbow trout macrophages synthesize lipoxins as major lipoxygenase products entirely from endogenous fatty acids. High-performance liquid chromatographic analysis of the supernatants from macrophages challenged with calcium ionophore A23187 revealed a range of lipoxygenase products including mono-hydroxy fatty acids, leukotrienes B4 and B5 and four major peaks with retention times and UV spectra characteristic of lipoxins (lambda max 302 nm). Cochromatography with authentic standards, UV spectroscopy and radiolabeling with [14C]arachidonate and eicosapentaenoate allowed tentative identification of the two largest peaks as lipoxin A4 and A5.

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Synthesis of leukotriene B and other conjugated triene lipoxygenase products by blood cells of the rainbow trout, Salmo gairdneri.

Stimulation of whole blood from rainbow trout with the calcium ionophore, A23187 (20 microM), produced leukotrienes B4 and B5 at concentrations in the range 22-30 ng.ml-1 and 8-24 ng.ml-1, respectively. Their identification and quantification was achieved using reverse-phase high-performance liquid chromatography, combined capillary column gas chromatography-electron capture chemical ionization mass spectrometry and ultraviolet spectroscopy. A number of other lipoxygenase products were also detected, but only partially analysed. The fatty acid composition of the leucocytes, which are presumed to be the site of leukotriene synthesis, was determined by thin-layer and gas-liquid chromatography to enable a comparison of the relative levels of the polyunsaturated fatty acids, which act as substrates for the synthesis of these lipoxygenase products. Arachidonic (20:4(n - 6)), eicosapentaenoic (20:5(n - 3)) and docosahexaenoic (22:6(n - 3)) acids represented approx. 6, 5 and 40%, respectively, of the total fatty acid content.

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