Patrick B. Costello

In vivo activation of an ω-6 oxygenase in human skin

To test the hypothesis that an epidermal fatty acid oxygenase is activated in vivo under physiologic conditions, surface lipids from normal human skin were analyzed for oxygenase products. With high-performance liquid chromatography on reversed-phase and straight-phase chiral columns and gas-liquid chromatography/mass spectrometry, these lipids were found to contain free 13-hydroxyoctadeca-9Z,11E-dienoic acid and 9-hydroxyoctadeca-10E,12Z-dienoic acid. The 13-hydroxyoctadecadienoic acid was present as a stereoisomeric mixture, with an average S/R ratio of 2.2, and exceeded the concentration of 9-hydroxyoctadecadienoic acid by a factor of 2. These observations and others indicate that the 13-hydroxyoctadecadienoic acid was derived mostly from an ω-6 oxygenase (probably 15-lipoxygenase) which is activated in vivo in normal skin.

Saturability of esterification pathways of major monohydroxyeicosatetraenoic acids in rat basophilic leukemia cells

The principal monohydroxyeicosatetraenoic acids (HETEs), 5-, 12-, and 15-HETE, which can be produced by rat basophilic leukemia (RBL-1) cells, are also esterified by these cells. Exogenously added 5-, 12-, and 15-HETE were rapidly incorporated as esters in RBL cells, reaching plateau levels within 25 min. In incubations in culture medium with protein added, all three HETEs were essentially completely metabolized within 24 h. 5-HETE was esterified more rapidly and to a greater extent than 12-HETE or 15-HETE when these were incubated together with RBL cells, indicating some degree of selectivity in the esterification pathways. When arachidonic acid (AA) was incubated in increasing concentrations with constant concentrations of 15-HETE and RBL cells, the free 15-HETE concentration increased and esterified 15-HETE concentration decreased markedly at AA: 15-HETE molar ratios above 9. 15-HETE esterification in RBL cells was also markedly inhibited by the polyunsaturated fatty acids, eicosatetraynoic and eicosapentanoic acids, but not by oleic or linoleic acids. In separate experiments with unlabeled and radiolabeled substrates, the extent of incorporation of esterified HETE in RBL cells decreased at higher concentrations of 15-HETE and AA, which showed that the pathway was saturable. The shapes of the curves for these fatty acid inhibitors suggest a concentration-dependent two compartment pathway of esterification. These data indicate that the HETEs and other 20 carbon fatty acid substrates probably compete for activity of a specific arachidonyl-CoA synthetase, which is the first and rate-limiting step for esterification of arachidonic acid by many human cells. Esterified 15-HETE was found to be predominantly in the phosphatidylethanolamine fraction of RBL cell lipids.

Lipoxygenase products in inflammatory synovial fluids and other exudates.

Forty six synovial fluid samples from 42 patients with inflammatory joint disease were analysed by reversed phase high performance liquid chromatography to determine 5-lipoxygenase products, specifically dihydroxyeicosatetraenoic acids (diHETEs). Twenty eight per cent of the fluids which were assayed had one or more products of 5-lipoxygenase activation. Seven fluids contained leukotriene B4 (0.1-28.1 ng/ml); three fluids had low concentrations of 20 carboxy/hydroxy-leukotriene B4 (0.01-0.05 ng/ml); three samples had leukotriene B4 isomers (1.5-2.4 ng/ml); and four fluids contained 5,15-diHETE (2.3-16.4 ng/ml). There was a poor correlation between synovial fluid white blood cell counts and evidence of 5-lipoxygenase activation. Several fluids contained unidentified compounds with spectra similar in shape to that of trienes, but the lambda max values of these unidentified compounds were different from those of known leukotrienes. A septic peritoneal exudate and a septic pleural fluid had concentrations of leukotriene B4 and leukotriene B4 isomers and metabolites in a range similar to those found in synovial fluids.

Case Report Dissected popliteal cyst: an unusual presentation of acute gout

A 46-year-old man with acute gout and a dissected popliteal cyst presented with clinical features which mimicked rheumatoid arthritis and thrombophlebitis. The clinical and laboratory diagnosis of the case are presented and similar cases previously reported are briefly reviewed.

Prevalence of occult inflammatory bowel disease in ankylosing spondylitis.

Fifty-five patients with ankylosing spondylitis and 16 control patients matched for sex and age were examined for evidence of occult inflammatory bowel disease. In all patients evaluation included history and physical examination, barium enema, sigmoidoscopy, and rectal biopsy. The results of this study suggest that there is no increased prevalence of occult inflammatory bowel disease in patients with ankylosing spondylitis.

Cytotoxicity of fatty acid oxygenase activation in rat basophilic leukemia cells

Apart from the generation of potent inflammatory mediators, the effects of fatty acid oxygenase activation, per se, on the host cell have not been well-delineated. Fatty acid oxygenases were activated in rat basophilic leukemia cells (RBL-1) by incubating them for 2-4 hr with 33-300 microM of arachidonic acid (AA) or linoleic acid (LA). As a control, the cells were incubated with one of two analogs of these fatty acids which are not oxygenase substrates: eicosatetraynoic acid or linoelaidic acid. Effects of oxygenase activation on cell viability were monitored by an assay for mitochondrial function. Cytotoxicity occurred in incubations with exogenous AA or LA in direct proportion to the substrate concentration but was not found in the control incubations or in incubations with the principal monohydroxylated AA products, 5-, 15-, and 12-HETE. Nordihydroguaiaretic acid (80 microM) and alpha-tocopherol (100 microM) significantly decreased the cell death observed during incubations with AA or LA. It is concluded that extensive oxygenase activation can result in cell death from intermediates produced proximal to the stable monohydroxylated derivatives.