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E F Ellis

Publications and source records attributed to E F Ellis.

At least 91 records · Page 5Linked to original sources

Lipoxygenase metabolism of arachidonic acid in brain.

When blood-free mouse brain slices were incubated with exogenous radiolabeled arachidonic acid, gas chromatography/mass spectrometry confirmed that the major radioactive lipoxygenase enzyme product of arachidonic acid was 12-hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE), with lesser amounts of 5-hydroxy-5,6,8,11,14-eicosatetraenoic acid and 15-hydroxy-5,8,11,13-eicosatetraenoic acid. When 12-[2H]HETE was used to measure endogenous 12-HETE in brain tissue frozen with liquid nitrogen, the level of 12-HETE was 41 +/- 6 ng/g of wet weight tissue. This frozen tissue level was not due to the presence of blood. When brain slices were incubated in vitro for 20 min, the 12-HETE level increased to 964 +/- 35 ng/g of wet weight tissue. Elimination of residual intravascular blood before tissue incubation reduced the brain slice 12-HETE concentration by one-half.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗

Reduction in cerebral arteriolar oxygen consumption by arachidonate.

The oxygen consumption of cerebral arterioles from anesthetized cats was measured using the Cartesian diver microrespirometer following in vitro incubation with 200 micrograms/ml of arachidonate or 50 micrograms/ml of 15-hydroperoxy-eicosatetraenoic acid (15-HPETE). Both agents depressed oxygen consumption severely. This effect was inhibited completely by a combination of superoxide dismutase (SOD) and catalase, indicating that it is mediated by oxygen radicals. Similar depression of oxygen consumption was observed during incubation of the vessels with xanthine oxidase and acetaldehyde as substrate. This enzymic system is known to generate superoxide and hydrogen peroxide. The effect of xanthine oxidase was also partially inhibited by SOD and catalase. The effect of arachidonate was partially inhibited by cyclooxygenase inhibitors. The effect of lipoxygenase inhibitors could not be adequately tested because they depressed oxygen consumption by themselves. Prostaglandins H2 and E2 had no effect on arteriolar oxygen consumption. The results show that arachidonate and 15-HPETE in high concentration depress cerebral arteriolar oxygen consumption via an oxygen radical-mediated mechanism. Furthermore, the radical is generated in the vessel wall and does not require either the brain parenchyma or the formed elements of the blood or the meninges for its production.

Animals↗

Appearance of superoxide anion radical in cerebral extracellular space during increased prostaglandin synthesis in cats.

When increased prostaglandin synthesis was induced in anesthetized cats equipped with cranial windows by topical application of arachidonate (200 micrograms/ml) or bradykinin (20 micrograms/ml), there was reduction of nitroblue tetrazolium, resulting in deposition of the reduced insoluble form of this dye on the brain surface. The amount of reduced nitroblue tetrazolium deposited on the brain surface was measured spectrophotometrically after fixation of the brain by perfusion with aldehydes to eliminate interference from hemoglobin. Topical application of 56 U/ml superoxide dismutase or 20 micrograms/ml indomethacin inhibited nitroblue tetrazolium reduction by 76.5%-82.5% and by 78%-85.5%, respectively. These results show that most of the nitroblue tetrazolium reduction was accounted for by superoxide anion radical generated in the course of arachidonate metabolism via the cyclooxygenase pathway. No superoxide production could be detected in the absence of arachidonate or bradykinin. Histological examination showed no evidence of parenchymal cellular damage or vascular damage and no accumulation of leukocytes. Pronounced leukocyte accumulation occurred 24 hours after topical arachidonate in rabbits with chronically implanted cranial windows. Superoxide appearance was reduced severely by 4,4'-diisothiocyano-2,2'-stilbene disulfonate and phenylglyoxal, two specific inhibitors of the anion channel. The most likely explanation for these findings is that increased metabolism of exogenous or endogenous arachidonate via cyclooxygenase results in the appearance of superoxide anion radical in cerebral extracellular space. Superoxide crosses the membrane of undamaged cells via the anion channel.

Animals↗

Evidence for a possible role of the brain kallikrein-kinin system in the modulation of the cerebral circulation.

Experiments by others have shown that exogenous bradykinin dilates cerebral arterioles and that the brain contains kininogen and kallikrein, the latter being the enzyme which converts kininogen to bradykinin. The objective of these experiments was to determine if bradykinin produced from endogenous brain kininogen can affect the cerebral microcirculation. Rabbit pial arteriolar diameter was measured with a microscope using the closed cranial window technique. Topical application of bradykinin (10(-8)-10(-5) M) induced a dose-dependent vasodilation (8-46%) which was completely inhibited by the cyclooxygenase enzyme inhibitors indomethacin and meclofenamic acid. Topical application of 1 U of tissue kallikrein per milliliter of artificial cerebrospinal fluid induced 43% dilation, which could be prevented by local treatment with indomethacin or the proteinase inhibitor aprotinin. The action of aprotinin and indomethacin was specific, since aprotinin did not affect the dilation produced by bradykinin, and indomethacin did not affect dilation produced by adenosine. A second application of kallikrein had no effect on cerebral diameter, yet the arterioles still responded normally to exogenous bradykinin, indicating that the first application of kallikrein depleted brain kininogen. We suggest that activation of brain kallikrein and subsequent formation of kinin from brain kininogen may be important in modulation of cerebral blood flow or generation of cerebral edema.

Animals↗

The effect of superoxide dismutase and catalase on metabolism of 3H-arachidonic acid by washed platelets.

Previous experiments have suggested that superoxide dismutase (SOD) and catalase (CAT) may inhibit prostaglandin synthesis. The purpose of this study was to determine if these free radical scavengers can alter the metabolism of free arachidonic acid (AA) by the cyclooxygenase and lipoxygenase enzyme systems in platelets. In control experiments washed platelets were incubated with 3H-AA for 5 minutes, extracted and the products separated by reverse phase high pressure liquid chromatography (HPLC). In normal intact platelets 13.5 +/- 0.6% of the radioactivity was found in TxB2, 16.3 +/- 1.4% in HHT, 61.3 +/- 1.1% in 12-HETE and 9.0 +/- 1.0% was unconverted AA. Pre-incubating the platelets for 1 minute with 10 micrograms/ml SOD or CAT or 10 micrograms/ml SOD plus 10 micrograms/ml CAT did not inhibit AA conversion or alter the percent product distribution. Similarly, SOD and CAT had no effect on AA metabolism in broken cells. However, as expected, pretreating platelets with indomethacin blocked TxB2 and HHT formation (P less than .0001). We conclude that SOD and CAT do not inhibit cyclooxygenase or lipoxygenase metabolism of free AA in platelets.

Animals↗

Effects of 15-hydroperoxy-eicosatetraenoic acid (15-HPETE) on cerebral arterioles of cats.

The effects of topical application of 15-hydroperoxy-eicosatetraenoic acid (15-HPETE, 200 micrograms/ml) on cerebral arterioles were studied in anesthetized cats equipped with cranial windows. 15-HPETE induced arteriolar dilation during application, sustained dilation 1 h after washout, and reduced responsiveness to the vasoconstrictive effects of hypocapnia. Electron microscopy of cerebral arterioles disclosed discrete endothelial lesions and focal morphological abnormalities of the vascular smooth muscle. Topical application of superoxide dismutase or catalase or the combination of the two inhibited the functional and morphological abnormalities induced by 15-HPETE. The results show that the vascular effects of 15-HPETE are mediated by superoxide anion radical and hydrogen peroxide or by other radicals derived from them, such as the hydroxyl radical. The results, together with earlier findings, support the view that the oxygen radicals responsible for these cerebral vascular effects are generated via the prostaglandin hydroperoxidase reaction.

Animals↗

Some properties of mouse platelets.

Mouse platelets were aggregated by arachidonate, thrombin, collagen and ADP. In general they were, like rat platelets, more aggregative in heparinized PRP than in citrated (3.8%) PRP. Mouse platelets underwent the release reaction when aggregated by arachidonate, collagen and thrombin, but not when stimulated by ADP. The aggregation of the platelets to arachidonate was inhibited by cyclooxygenase inhibitors and by prostacyclin. Studies with tritiated arachidonate showed that mouse platelets possess the lipoxygenase and cyclooxygenase pathways found in other mammalian platelets and produce thromboxane and 12-HETE. The mouse provides a convenient model for the study of many conditions known to affect platelet aggregation. The similarity of mouse platelets to the platelets of other mammals together with the ability to study large numbers of animals at low cost, should encourage further use of mouse platelets.

Adenosine Diphosphate↗

Conversion of arachidonic acid to cyclooxygenase and lipoxygenase products, and incorporation into phospholipids in the mouse neuroblastoma clone, Neuro-2A.

Arachidonic acid (AA) incorporation into phospholipids and cyclooxygenase and lipoxygenase mediated metabolism of arachidonic acid were studied in homogenized and intact Neuro-2A cells. When 3H8-AA was added to homogenized cells and incubated 20 minutes, 39% of the label was converted to prostaglandins (PGs), 10% to hydroxy-eicosatetraenoic acid (HETE) and 26% was incorporated into phospholipids. PGE2 and PGF2a were the major PGs produced. Synthesis of PGs was blocked by 10 microM indomethacin and synthesis of PGs and HETE was blocked by 10 microM eicosatetraynoic acid (ETYA). The cell homogenate produced the 13,14-dihydro-15-keto metabolites of PGE2 and PGF2a from 3H8-AA and also converted exogenous 3H7-PGE2 and 3H8-PGF2a to metabolites. When intact cells were labeled for 24 hours with 14C1-AA and the cells and media then analyzed, 75% of the radioactivity was incorporated into cellular phospholipids, 0.8% was converted to PGs and metabolites and 0.7% converted to HETE. Cells prelabeled for 24 hours were washed and incubated for 30 minutes in fatty acid free media. There was a 23% release of AA from phospholipids. One-fifth of the released AA was converted to HETE. PG synthesis in the intact resting cells was low. In summary, the Neuro-2A cell provides a good model system for studying arachidonic acid metabolism and incorporation into phospholipids in cells of neuronal origin.

Animals↗

Theophylline.

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Adolescent↗

The effect of PGF2 alpha on in vivo cerebral arteriolar diameter in cats and rats.

We compared the effect of topical application of PGF2 alpha on cerebral arterioles in cats and rats equipped with an acutely implanted cranial window. Arterial diameter was measured using a microscope and image splitting device. PGF2 alpha in a concentration ranging from 10(-7) to 10(-5) M had no effect on large (greater than or equal to 100 microns) or small (less than 100 microns) cat pial arterioles, but induced a dose dependent constriction of rat pial arterioles with a maximum constriction to 76% of control diameter. Dilation of cat large cerebral arterioles by topically applied PGE2 was not affected by simultaneous application of PGF2 alpha and PGE2 induced dilation of small arterioles was decreased 3% by PGF2 alpha. While we and others have previously shown that both cat and rat brain can synthesize PGF2 alpha, it appears that PGF2 alpha is not likely to normally be a major modulator of cerebral arteriolar resistance in all species.

Animals↗

Asthma in childhood.

Asthma is defined as an obstructive disease of the pulmonary airways resulting from spasm of airway muscle, increased mucus secretion, and inflammation. The airways of asthmatic individuals are hyperresponsive to a variety of stimuli including cold air, atmospheric irritants, pharmacologically active chemicals, various drugs, and hyperventilation. The fundamental abnormality underlying the hyperresponsiveness appears to be genetically determined; two theories explaining the abnormality have received the most attention. One theory suggests that asthma is due to abnormal beta-adrenergic receptor-adenylate cyclase function with decreased adrenergic responsiveness. An alternate theory proposes that increased cholinergic activity in the airway is the fundamental defect in the disease. The true prevalence of asthma has been difficult to determine owing to uncertainties regarding the definition of the disease. Prevalence in various populations of children ranged from 1.37% to 11.4% or higher. Most studies report a preponderance of asthma in boys over girls, with ratios varying from 1.3:1 to 3.3:1. Risk factors for the disease include a history of atopy, acute lower respiratory tract disease, parental cigarette smoking, and bronchiolitis or croup. The spectrum of asthma is that of an illness beginning early in life and persisting, in some cases, through adulthood. Signs of the disease may be apparent in the first 2 yr of life and are often associated with viral respiratory infections. Disproportionate narrowing of peripheral airways and decreased static elastic recoil properties of the lung predispose infants and young children to asthma. During midchildhood there is a tendency toward improvement, with continued improvement during adolescence. The goal of management of the child with asthma is to reduce symptoms sufficiently so that the child can regularly attend school, engage in play activities, and sleep through the night uninterrupted, while avoiding unacceptable levels of adverse drug effects. Nonpharmacologic management includes both avoidance of environmental irritants and behavioral approaches to overcome emotional precipitants that lead to attacks. Pharmacologic treatment includes the use of four classes of drugs: (1) adrenergics, (2) theophylline, (3) cromolyn, and (4) corticosteroids. The prognosis of asthma in childhood is good. Although airway activity may remain abnormal for indefinite periods of time, most children reach a state where they are virtually free of symptoms.

Adolescent↗

Effect of fasting on production from exogenous arachidonic acid of thromboxane and 12-hydroxy 5, 8, 10, 14-Eicosatetraenoic acid (12-HETE) by washed platelets of normal and streptozotocin diabetic mice.

Washed mouse platelets produced thromboxane B2 (TXB2) and 12-hydroxy 5, 8, 10, 14-Eicosatetraenoic acid (12-HETE) in vitro from tritiated arachidonic acid. When diabetes was induced by injecting streptozotocin i.p 200 mg/kg the TXB2 production fell significantly and 12-HETE production significantly increased compared with that of control animals. Eighteen hours of food deprivation caused significant reductions in TXB2 and 12-HETE production in both the control and diabetic groups.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗