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Biomedical subjects

J R Hoult

Publications and source records attributed to J R Hoult.

At least 19 recordsLinked to original sources
PubMed1992

Interactions of a series of coumarins with reactive oxygen species. Scavenging of superoxide, hypochlorous acid and hydroxyl radicals.

Sixteen plant-derived or synthetic coumarins with various hydroxyl and other substitutions were tested for their ability to inhibit lipid peroxidation and to scavenge hydroxyl radicals, superoxide radicals and hypochlorous acid. Seven unsubstituted or monosubstituted coumarins were essentially inactive in all tests except for ability to scavenge OH with rate constants approximately greater than 1 x 10(9) M-1. sec-1. Of the remaining nine, six containing dihydroxy substitutions were effective inhibitors of Fe3+-ascorbate-dependent microsomal lipid peroxidation (IC50 less than 20 microM), with ortho-dihydroxy + one additional substitution optimal (IC50 less than 10 microM). ortho-Dihydroxylated coumarins were pro-oxidant (enhanced OH generation) in the Fe3+-EDTA-H2O2 deoxyribose system but decreased OH' generation in the Fe3+-ascorbate-H2O2 deoxyribose system, indicating that these compounds can both chelate iron ions and also readily donate electrons for redox cycling of Fe3+. The meta-dihydroxycoumarin did not show this behaviour, but was an effective scavenger of hypochlorous acid, a property shared by only one other compound. Several other coumarins with one or more hydroxyl substituents were also capable of effectively removing superoxide anions (IC50 3.7-72 microM), although some could not be quantified due to direct rapid reduction of cytochrome c. We conclude that several compounds, notably 5,7-dihydroxy-4-methylcoumarin, possess beneficial biochemical profiles of interest in relation to pathophysiological processes dependent upon reactive oxygen species.

Animals↗
PubMed1992

Inhibition of 5-lipoxygenase and cyclo-oxygenase in leukocytes by feverfew. Involvement of sesquiterpene lactones and other components.

Leaves or infusions of feverfew, Tanacetum parthenium, have long been used as a folk remedy for fever, arthritis and migraine, and derived products are widely available in U.K. health food shops. Previous reports have suggested interactions with arachidonate metabolism. Crude chloroform extracts of fresh feverfew leaves (rich in sesquiterpene lactones) and of commercially available powdered leaves (lactone-free) produced dose-dependent inhibition of the generation of thromboxane B2 (TXB2) and leukotriene B4 (LTB4) by ionophore- and chemoattractant-stimulated rat peritoneal leukocytes and human polymorphonuclear leukocytes. Approximate IC50 values were in the range 5-50 micrograms/mL, and inhibition of TXB2 and LTB4 occurred in parallel. Isolated lactones (parthenolide, epoxyartemorin) treated with cysteine (to neutralize reactive alpha-methylene butyrolactone functions of the sesquiterpenes). Inhibition of eicosanoid generation appeared to be irreversible but not time-dependent. We conclude that feverfew contains a complex mixture of sesquiterpene lactone and non-sesquiterpene lactone inhibitors of eicosanoid synthesis of high potency, and that these biochemical actions may be relevant to the claimed therapeutic actions of the herb.

Animals↗
PubMed1992

Eicosanoid synthesis and Helicobacter pylori associated gastritis: increase in leukotriene C4 generation associated with H. pylori colonization.

The importance of pro-inflammatory leukotriene C4 in Helicobacter pylori (H. pylori) associated gastritis in man is unknown. Fresh gastric biopsy specimens from 28 dyspeptic patients were obtained: 10 showed normal antral histology with no evidence of H. pylori, the remaining 18 patients exhibited histological gastritis and were H. pylori positive as assessed by histology, culture and urease test. Twelve of these 18 patients received 240 mg twice daily colloidal bismuth subcitrate for four weeks before re-endoscopy. Gastric biopsies from H. pylori positive patients were incubated under basal and Ca(2+)-ionophore mediated conditions: Radioimmunoassay analysis of the supernatant showed basal release of prostaglandin E2 and leukotriene C4 was slightly but not significantly elevated in H. pylori positive mucosa. However in H. pylori positive mucosa there was an 85% increase in leukotriene C4 synthesis when biopsies were incubated with ionophore, compared to only 13% increase in H. pylori negative mucosa (p less than 0.02). After eradication of H. pylori by colloidal bismuth subcitrate, there was a clearance of inflammatory cell infiltrate as assessed by histology and a significant reduction in ionophore-mediated leukotriene C4 formation compared with before treatment (p less than 0.02). These results suggest that H. pylori gastritis is associated with increased capacity to generate leukotriene C4, which may amplify the damaging effects of the bacteria on gastric mucosa.

Adult↗
PubMed1992

Feverfew extracts and parthenolide irreversibly inhibit vascular responses of the rabbit aorta.

Samples prepared from chloroform extracts of fresh leaves of feverfew (Tanacetum parthenium) strongly inhibited responses of rabbit aortic rings to phenylephrine, 5-hydroxytryptamine, thromboxane mimetic U46619 (9,11-dideoxy-11 alpha,9 alpha-epoxy-methano-PGF2 alpha), and angiotensin II, but the inhibition to contractions induced by potassium depolarization was much less. The inhibition was concentration- and time-dependent, non-competitive, and irreversible, and also occurred in endothelium-denuded preparations. The feverfew extracts also caused a progressive loss of tone of pre-contracted aortic rings and appeared to impair the ability of acetylcholine to induce endothelium-dependent relaxations of the tissue. These effects were mimicked by a purified preparation of an alpha-methylenebutyrolactone, parthenolide, obtained from the extract. Our results demonstrate a nonspecific and potentially toxic response to feverfew on the vasculature.

Acetylcholine↗
PubMed1992

Eicosanoid synthesis in duodenal ulcer disease: decrease in leukotriene C4 by colloidal bismuth subcitrate.

The release of immunoreactive prostaglandin E2 (PGE2) and leukotriene C4 (LTC4) from antral and duodenal mucosal biopsy specimens taken from 20 patients with duodenal ulcer disease was measured by radioimmunoassay before and four weeks after treatment with colloidal bismuth subcitrate. Gastroscopic and histological examination showed complete ulcer healing in 15/18 patients and duodenal histology looked normal (n = 15) or improved (n = 3): two patients failed to attend for a second endoscopy. Analysis of the supernatant from incubations of biopsy tissue in vitro showed that unstimulated antral release of PGE2 was significantly more than that from the duodenal mucosa (p less than 0.05), whereas basal release of LTC4 was significantly lower from antral biopsy specimens (p less than 0.05). Subsequent incubation of specimens with calcium ionophore A23187 caused an increase in LTC4 but not in PGE2 generation. The ability of antral and duodenal mucosa to form ionophore mediated LTC4 in patients with duodenal ulcer disease was significantly greater (p less than 0.05; p less than 0.01 respectively) than that of normal gastroduodenal mucosa. After colloidal bismuth subcitrate treatment, basal synthesis of PGE2 was unchanged in duodenal and antral specimens. In contrast, basal duodenal LTC4 was reduced (p less than 0.05), and the capacity for ionophore mediated duodenal LTC4 formation was substantially and significantly reduced after treatment (p less than 0.001). These results indicate that after therapeutic healing of duodenal ulcer (accompanied by clearance of inflammatory cell infiltrate), there is a reduced ability of duodenal mucosa to generate proinflammatory peptidoleukotrienes.

Adult↗
PubMed1992

Peroxyl radical scavenging by a series of coumarins.

Sixteen plant-derived or synthetic coumarins with various hydroxyl and other substitutions were tested for their ability to scavenge alkylperoxyl radicals generated in the aqueous phase by the controlled thermolysis of 2,2'-azo-bis-(2-amidinopropane) dihydrochloride (ABAP). Protection by coumarins against inactivation of lysozyme by the radicals was assayed by measuring the loss of turbidity of suspensions of M. lysodeikticus. Ten of the coumarins were potent scavengers of aqueous peroxyl radicals with activities comparable to n-propyl gallate, desferrioxamine, ferrioxamine and trolox c, yielding IC50 values in the range 21 to 92 micromolar. The presence of 6,7-ortho-dihydroxy functions gave compounds of the greatest potency. Scavenging activity was unrelated to ability to chelate iron ions. The active coumarins are attractive candidates for evaluation as protective agents against disorders in which oxidative stress is implicated.

Amidines↗
PubMed1991

Inhibition of mammalian 5-lipoxygenase and cyclo-oxygenase by flavonoids and phenolic dietary additives. Relationship to antioxidant activity and to iron ion-reducing ability.

We investigated the ability of various plant flavonoids (a) to inhibit 5-lipoxygenase and cyclooxygenase activities in rat peritoneal leukocytes, (b) to inhibit lipid peroxidation in rat liver microsomes, and (c) to stimulate DNA degradation caused by the antibiotic bleomycin in the presence of ferric ions. These compounds were compared with a range of synthetic phenolic substances including carnosol, vanillin, vitamin E and its analogue trolox c. The flavonoids were potent inhibitors of non-enzymatic peroxidation in membranes but this was not significantly correlated with their ability to inhibit either pathway of eicosanoid synthesis, suggesting that their mode of inhibition of 5-lipoxygenase/cyclooxygenase is not simply due to interception of peroxyl radicals generated at the active site of the enzymes. Many of the flavonoids and other compounds (including carnosol, vitamin E and trolox c) stimulated Fe3+/bleomycin-dependent DNA degradation. Those flavonoids which stimulated DNA degradation at low concentrations but which inhibited it at higher concentrations ("biphasic" effect, possibly caused by changing relative contributions of ability to reduce ferric-bleomycin or to chelate iron ions from the bleomycin) were selective inhibitors of 5-lipoxygenase compared to cyclo-oxygenase. In contrast, those flavonoids that did not stimulate DNA degradation at all proved to be cyclo-oxygenase selective inhibitors. Compounds that increased Fe3+/bleomycin-dependent DNA damage up to a maintained plateau were non-selective inhibitors of both 5-lipoxygenase and cyclo-oxygenase. Thus, a combination of iron-chelating and iron ion-reducing properties appears to be required for selective 5-lipoxygenase inhibition by phenolic compounds. Carnosol, vitamin E and trolox c were also found to be 5-lipoxygenase inhibitors of varying potency, and all were less active as cyclo-oxygenase inhibitors.

Abietanes↗
PubMed1990

Flavonoids inhibit the oxidative modification of low density lipoproteins by macrophages.

Low density lipoproteins (LDL) can be oxidatively modified in vitro by macrophages and certain other cell types so that macrophages will take them up much faster. This process may be important in the formation of cholesterol-laden foam cells derived from macrophages in atherosclerotic lesions. In this study, we have shown that certain flavonoids, plant constituents found in the diet, are potent inhibitors of the modification of 125I-labelled LDL by macrophages, with IC50 values in the micromolar range (e.g. morin and fisetin 1 microM; quercetin and gossypetin 2 microM). The potencies of individual flavonoids in inhibiting LDL modification did not correlate with their previously determined potencies as inhibitors of 5-lipoxygenase and cyclo-oxygenase. The modification of LDL by macrophages exhibits a lag period of about 4-6 hr before enhanced uptake is detected. During this time, there is a rapid depletion in its content of alpha-tocopherol (an endogenous antioxidant found in lipoproteins) followed by a large increase in the level of hydroperoxides. The flavonoids conserved the alpha-tocopherol content of LDL and delayed the onset of detectable lipid peroxidation. Flavonoids also inhibited the cell-free oxidation of LDL mediated by CuSO4. These findings raise the possibility that flavonoids may protect LDL against oxidation in atherosclerotic lesions and may therefore be natural anti-atherosclerotic components of the diet, although this will depend to a large extent on their pharmacokinetics.

Copper↗
PubMed1990

Alpha-tocopherol consumption during low-density-lipoprotein oxidation.

1. The kinetics of the depletion of alpha-tocopherol in human low-density lipoprotein (LDL) were measured during macrophage-mediated and cell-free oxidation. The formation of oxidatively modified, high-uptake species of LDL in these systems was not detectable until all of the endogenous alpha-tocopherol had been consumed. 2. Supplementation of the alpha-tocopherol content of LDL by loading in vivo extended the duration of the lag period during which no detectable oxidative modification occurred. 3. The addition of a flavonoid (morin) prevented both alpha-tocopherol consumption and oxidative modification of LDL. 4. The alpha-tocopherol contents of LDLs from a range of individual donors could not be used to predict their relative resistance to oxidation, indicating that other endogenous antioxidants may also be present, and quantitatively significant, in human LDL.

Animals↗
PubMed1990

Differential regulation of 5-lipoxygenase and cyclo-oxygenase pathways of arachidonate metabolism in rat peritoneal leukocytes.

1. Simultaneous activation of the 5-lipoxygenase and cyclo-oxygenase pathways of arachidonate metabolism in rat peritoneal mixed leukocytes in response to A23187, chemoattractant N-formyl-methionine-leucine-phenylalanine (FMLP) and arachidonic acid (AA) was studied by radioimmunoassay of leukotriene B4 (LTB4) and thromboxane B2 (TXB2) respectively. 2. FMLP and AA preferentially activated cyclo-oxygenase and A23187 preferentially activated 5-lipoxygenase. Release of TXB2 preceded that of LTB4. A threshold amount of A23187 enhanced FMLP-and AA-induced LTB4 production but not that of TXB2. 3. Phorbol myristate acetate (PMA) abolished LTB4 generation in response to FMLP with much less effect on TXB2, but did not inhibit the formation of either eicosanoid caused by A23187 or AA. Instead, PMA caused a dose-dependent but modest stimulation of TXB2 generation either on its own or when added with A23187 or AA. 4. These results show that the 5-lipoxygenase and cyclo-oxygenase pathways in rat peritoneal leukocytes are regulated differently and that functional compartmentalisation of the stimulus-generation sequence operates in these cells.

Animals↗
PubMed1989

Antioxidant and pro-oxidant actions of the plant phenolics quercetin, gossypol and myricetin. Effects on lipid peroxidation, hydroxyl radical generation and bleomycin-dependent damage to DNA.

The plant-derived phenolic compounds gossypol, quercetin and myricetin are powerful inhibitors of iron-induced lipid peroxidation in rat liver microsomes, under all five experimental conditions tested and at low micromolar concentrations (IC50 less than or equal to 1.5 microM). However, they greatly accelerate the generation of hydroxyl radicals (.OH) from H2O2 in the presence of Fe3+-EDTA at pH 7.4, as measured by the deoxyribose assay. At 100 microM, the three phenolic compounds enhanced .OH formation up to eight-fold. The hydroxyl radical generation was inhibited by catalase and superoxide dismutase, suggesting a mechanism in which the phenols oxidize to produce superoxide radical, which then assists .OH generation from H2O2 in the presence of Fe3+-EDTA. At concentrations up to 75 microM, quercetin and myricetin also accelerate bleomycin-dependent DNA damage in the presence of Fe3+, possibly by reducing the Fe3+-bleomycin-DNA complex to the Fe2+ form. Hence these naturally-occurring substances can have pro-oxidant effects under some reaction conditions and cannot be classified simplistically as "antioxidants".

Animals↗
PubMed1989

Platelet activating factor is a potent colonic secretagogue with actions independent of specific PAF receptors.

Several candidate mediators of acute inflammation such as E-type prostaglandins, histamine and bradykinin are potent pro-diarrhoeal colonic secretagogues. They act to increase serosal to mucosal transport of chloride and passive water efflux. We investigated the effects of platelet activating factor (PAF) on muscle-stripped rat colon, measuring transepithelial potential difference (p.d.) and, under voltage clamp conditions, short circuit current (Isc). PAF induced dose-dependent increases in p.d. and Isc, with an approximate EC50 of 1.5 X 10(-10) M; similar concentrations of lyso-PAF had a much smaller but discernible effect. PAF and lyso-PAF both displayed 'sidedness' with serosal application effective and mucosal application ineffective. Inhibitor studies suggest that chloride is the principal ion carrier, but the specific PAF receptor antagonists kadsurenone, L652731, CV 3988 and WEB 2086 did not block the response. Unlike bradykinin, PAF did not cause the release of PGE2 into the serosal bathing fluid, and its action was not attenuated by the cyclo-oxygenase inhibitors piroxicam, mefenamic acid or flurbiprofen. We conclude that PAF has a powerful pro-diarrhoeal secretory action on colonic epithelium which is not mediated by the previously defined PAF receptor(s) and is independent of prostanoid generation.

Animals↗
PubMed1989

Effects of desferrioxamine on eicosanoid production in two intact cell systems.

Desferrioxamine is an iron-chelating agent used in the treatment of iron overload. It is a powerful inhibitor of iron-dependent radical reactions and has been used to test for the involvement of such reactions in animal models of human disease. Recent proposals that desferrioxamine inhibits lipoxygenase and stimulates cyclo-oxygenase were examined by testing its action upon eicosanoid production in rat caecal fragments and elicited rat peritoneal leukocytes. Under certain experimental conditions, higher concentrations of desferrioxamine (up to 0.5 mM) enhanced eicosanoid production in leukocytes. Desferrioxamine concentrations greater than or equal to 500 microM also stimulated cyclo-oxygenase activity in caecal fragments. Although these results show that high concentrations of desferrioxamine can augment eicosanoid production under certain circumstances, the effects are likely not to be significant therapeutically in view of the concentrations of desferrioxamine achieved during in vivo use in humans.

Animals↗
PubMed1988

Organ selective conversion of prostaglandin D2 to 9 alpha, 11 beta-prostaglandin F2 and its subsequent metabolism in rat, rabbit and guinea pig.

Cell-free 100,000 g supernatants from liver, kidney, lung and caecum of rat, rabbit and guinea-pig were compared for their ability to transform prostaglandins F2 alpha, D2, E2 and 9 alpha, 11 beta-prostaglandin F2 (11epi-PGF 2 alpha) to metabolic products. Experiments utilized multitritiated substrate PGs, with assessment of biotransformation by TLC, HPLC and GC/MS. PGF2 alpha was converted via the sulphasalazine analogue-inhibitable NAD+-dependent 15-hydroxy-prostaglandin dehydrogenase pathway (15-PGDH), with high activity (greater than 5 pmol/min/mg protein) in all 12 systems except rat and rabbit liver (e.g. guinea-pig kidney and rat caecum both 64 pmol/min/mg; rat liver 0.3 pmol/min/mg), forming 15-keto and 13,14-dihydro-15-keto metabolites as determined by TLC, HPLC and GC/MS. Prostaglandin D2 was not transformed in similar fashion in NAD+- or NADP+-supplemented incubations in any of the 12 cytosolic systems. However, PGD2 was converted to a single product identified by TLC, HPLC and GC/MS as 9 alpha, 11 beta-PGF2 in certain of the systems when supplemented with an NADPH regenerating system, with high activity in guinea-pig kidney (55.0 pmol/min/mg), guinea-pig liver (27.5 pmol/min/mg) and rabbit liver (13.7 pmol/min/mg) and less than 5 pmol/min/mg in 8 of the remaining 9 systems. This stereospecific 11-ketoreductase of rabbit and guinea-pig liver was stable to 10 min heating at 50 degrees, dialysis, storage at -20 degrees and repeated freeze/thawing but was not inhibited by sulphasalazine analogues. The 11-ketoreductase had a markedly different tissue profile from PGE2 9-ketoreductase, which was shown to convert PGE2 stereospecifically to 9 alpha, 11 alpha-prostaglandin F2 (PGF2 alpha) and was present at highest activity in rabbit liver and kidney. Evidence was obtained that 9 alpha, 11 beta-PGF2 was actively transformed by the sulphasalazine-inhitable 15-PGDH pathway at approximately one third of the rate of PGF2 alpha with high activity in several cytosolic systems (e.g. rat caecum, guinea-pig liver and kidney), suggesting that further transformation in vivo of this biologically active product of PGD2 metabolism could be initiated by this route.

Animals↗
PubMed1988

Oxidants, inflammation, and anti-inflammatory drugs.

Species such as superoxide radical (O2-), hydrogen peroxide (H2O2), hydroxyl radical (.OH), and hypochlorous acid (HOCl) can be formed in vivo, e.g., by activated phagocytic cells. Generation of .OH from H2O2 in vivo usually involves iron-dependent reactions. Good evidence exists for increased generation of oxidants in vivo in patients with active rheumatoid disease, but the contribution of these oxidants to the disease process is still uncertain. The likelihood that anti-inflammatory drugs used in the treatment of arthritis could act by scavenging oxidants or preventing their formation is discussed.

Anti-Inflammatory Agents↗
PubMed1988

Secretory effects of kinins on colonic epithelium in relation to prostaglandins released from cells of the lamina propria.

1. Sheets of muscle-stripped rat and rabbit colon with epithelium intact or removed were mounted in Ussing-type chambers for recording of transepithelial p.d., resistance and short circuit current (Isc), and measurement by radioimmunoassay (RIA) of the release of prostaglandins into serosal and mucosal bathing solutions. 2. In epithelial-intact preparations prostaglandin E2 (PGE2), PGE1, PGF2 alpha, U46619 and prostacyclin (10(-7)-10(-6) M) caused increases in Isc and transepithelial p.d., in (approximate) descending order of potency. Epithelial-removed preparations did not exhibit any transepithelial p.d. 3. In epithelial-intact preparations, lysyl-bradykinin (LBk) applied serosally but not mucosally caused increased p.d. and release of PGE2 (and to a lesser extent other prostaglandins) into serosal but not mucosal bathing solutions. In epithelial-removed tissues, responsiveness to LBk was maintained, but it did not exhibit 'sidedness', i.e. LBk was effective when applied on either side and PGE2 release occurred into both compartments. 4. Indomethacin and other non steroidal anti-inflammatory drugs (NSAIDs) abolished the LBk-induced p.d. and reduced PGE2 release if applied serosally but not mucosally in epithelial-intact preparations. In epithelial-removed tissues, indomethacin added to either side abolished prostaglandin release into both compartments. 5. Calcium removal from serosal but not mucosal bathing solution (Ca2+-free EGTA Krebs) abolished p.d. generation by LBk in epithelial-intact preparations, and reduced PGE2 release in rabbit but not rat colon. Similarly, in epithelial-removed preparations, calcium removal did not affect kinin-induced PGE2 generation in rat but strongly attenuated it in rabbit colon. 6. We conclude that (i) kinins activate the arachidonate cascade principally by interactions with cells in the subepithelial (lamina propria) layer, rather than with the epithelial cells themselves, (ii) PGE2 contributes substantially to the kinin-induced increase of transepithelial p.d. as a messenger released from kinin-responsive subepithelial cells and acting on the basolateral pole of the epithelial cells, (iii) the apparent sidedness of colonic epithelium in terms of responses to kinins, NSAIDs and calcium removal is due to the barrier properties of the epithelial cell layer, and (iv) there are differences in calcium sequestration and apparent calcium dependence of prostaglandin biosynthesis between rat and rabbit colonic subepithelial cells.

Animals↗