Biomedical subjects

J B German

Publications and source records attributed to J B German.

82 records · Page 5Linked to original sources

PubMed1990

Phospholipid fatty acid composition of various mouse tissues after feeding alpha-linolenate (18:3n-3) or eicosatrienoate (20:3n-3).

The selective incorporation of dietary alpha-linolenate (18:3n-3) and its elongation product, eicosatrienoate (20:3n-3), into various phospholipids (PL) of mouse liver, spleen, kidney, and heart, was examined in a two-week feeding trial by assessing mol % changes in associated fatty acids. Mice were fed fat-free AIN 76A diets modified with either 2 wt% safflower oil (control); 1% safflower and 1% linolenate; or 1% safflower and 1% eicosatrienoate. After linolenate or eicosatrienoate feeding, 20:4n-6 was reduced by 36-50% in liver phosphatidylcholine (PC) and in liver and spleen phosphatidylethanolamine (PE). Linolenate was minimally incorporated into PL, but was desaturated and elongated to 20:5n-3, 22:5n-3, and 22:6n-3, with notable differences in the quantity of these n-3 derivatives associated with different tissues and PL. Eicosatrienoate was uniquely incorporated into the cardiolipin (CL) pool of all organs. There was also considerable retroconversion of 20:3n-3 to 18:3n-3 (PC,PE). Dietary eicosatrienoate may therefore affect metabolism in diverse ways--20:3n-3, which is retroconverted to 18:3n-3, may provide substrate for 20:5n-3 and 22:6n-3 syntheses, whereas intact 20:3n-3 may be incorporated into the CL pool. Acyl modifications of CL are known to affect the activity of key innermitochondrial enzymes, such as cytochrome c oxidase.

8,11,14-Eicosatrienoic Acid↗

PubMed1990

Oxidant stress inhibits the endogenous production of lipoxygenase metabolites in rat lungs and fish gills.

Hydroperoxides are potent initiators of lipid peroxidation in vivo. Acyl hydroperoxides may also regulate various aspects of lipid metabolism. In this study we investigated the regulation of the endogenous 12 lipoxygenase in trout gill and rat lung, a prominent acyl hydroperoxide catalyst in these tissues. Initial experiments revealed that the enzyme from trout gill was activated by hydroperoxides at low levels and inactivated by the same hydroperoxides at high levels. Homogenization of these tissues resulted in the production of a predominant metabolite class from released endogenous polyunsaturated fatty acids, the 12 lipoxygenase products. In rat lung, arachidonic acid was the major polyunsaturated fatty acid released and 12 (S) HETE was the major metabolite. In trout gill 20:4, 20:5 n3, and 22:6 n3 were released and the 12(S), 12, and 14 hydroxy derivatives the corresponding metabolites. Computer simulations of the sensitivity of these enzymes to hydroperoxides predicted that exogenous oxidant stress would reduce significantly the production of HETEs. Tertiary butyl hydroperoxide was added to tissue homogenates and resulted in elimination of greater than 95% of the lipoxygenase activity. These results suggest that the lipoxygenase enzyme in lung and gill tissue is a major potential source for acyl hydroperoxides in vivo, but is also very sensitive to oxidant stresses including the acyl hydroperoxides themselves. This enzyme could thus be an important focus for oxidant injury in lungs.

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

PubMed1988

Docosahexaenoic acid and other dietary polyunsaturated fatty acids suppress leukotriene synthesis by mouse peritoneal macrophages.

The efficacy of individual omega-3 polyunsaturated fatty acids (PUFA) in altering eicosanoid synthesis in peritoneal macrophages was studied by feeding mice for 10 days a diet containing 2 wt % fat, which included 0.5 wt % ethyl esters of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) or linolenic acid (LNA). Upon stimulation with calcium ionophore A23187, macrophages from these animals produced significantly lower amounts of leukotriene C4, leukotriene B4 and 12-hydroxyeicosatetraenoic acid, prostaglandin E2 and 6-keto prostaglandin F1 alpha compared with those obtained from animals on the diets containing olive oil or safflower oil. The decrease in leukotriene synthesis was similar in the animals fed DHA, EPA or LNA diets. This depression of eicosanoids by DHA and EPA was associated with decreased levels of arachidonic acid (AA); however, LA that altered eicosanoids did not have the same effect on AA levels.

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

PubMed1988

Relative inhibitory potencies of flavonoids on 12-lipoxygenase of fish gill.

The relative efficacy of 10 flavonoid compounds and some common antioxidants in inhibiting 12-lipoxygenase of fish gill was determined. Lipoxygenase activity was measured by oxygen consumption with polarography and formation of hydroxy fatty acid using thin layer chromatography with autoradiography. Generally, the inhibition of 12-lipoxygenase by flavonoids was noncompetitive. The lipoxygenase product pattern did not change in the presence of flavonoids. Fisetin (3,3',4',7-tetrahydroxyflavone) and quercetin (3,3',4',5,7-pentahydroxyflavone) were the most potent inhibitors of 12-lipoxygenase, with IC50's of 0.25 and 0.4 microM respectively. These were compared with IC50's values of 5,1000 microM for BHA (t-butylhydroxyanisole) and BHT (t-butylhydroxytoluene) respectively. Possible inhibitory mechanisms and relationships between flavonoid structure and inhibitory potencies are discussed.

Animals↗

PubMed1988

The effect of dietary fish oils on eicosanoid biosynthesis in peritoneal macrophages is influenced by both dietary N-6 polyunsaturated fats and total dietary fat.

Recent research has implicated dietary fish oils in the reduction of eicosanoids formed from arachidonic acid and amelioration of chronic diseases such as coronary heart disease, atherosclerosis and inflammation. Feeding studies were conducted to determine if the efficacy of dietary n-3 polyunsaturated fatty acids (PUFA) from fish oils was influenced by the quantity of n-6 polyunsaturated fatty acids and the total level of fat in the diet. Groups of mice were fed diets composed of 5 and 20% total fat with varying proportions of linoleic acid as a source of n-6 PUFA. Menhaden oil as a source of n-3 PUFA was fed at two levels of n-6 at each level of total fat. Eicosanoid biosynthesis was stimulated and assayed in the mouse peritoneum using zymosan as an inflammatory stimulus. Production of LTE4 and PGE2 was enhanced by increasing n-6 PUFA in the diet at both levels of total fat. High dietary fat significantly suppressed leukotriene (LT) synthesis. Dietary menhaden oil reduced LTE4 and PGE2 synthesis at both levels of dietary n-6 in the low fat study. In animals on 20% dietary fat menhaden oil significantly reduced LT synthesis only at a relatively low dietary n-6 PUFA. On a high n-6 PUFA high fat diets, menhaden oil did not significant affect LTE4 synthesis in response to zymosan stimulation. The results suggest that the effectiveness of fish oils in reducing eicosanoids in response to specific stimulation is influenced by the level of n-6 and the total quantity of fat in the diet.

Animals↗

PubMed1987

Modulation of zymosan stimulated leukotriene release by dietary unsaturated fatty acids.

The quantity of leukotrienes produced in an inflammation model, the stimulated mouse peritoneum, was affected by dietary manipulation of tissue arachidonic acid, the immediate leukotriene (LT) precursor. Fifty ng of LTE4 was synthesized (after injection of zymosan) by the peritoneal cavity of mice maintained on olive oil as a dietary source of unsaturated fatty acids. Animals maintained on corn oil, exhibited significantly enhanced leukotriene biosynthesis upon stimulation by zymosan. Mice fed menhaden oil, a fat source rich in n-3 fatty acids produced 50% less leukotriene E4 than animals fed olive oil. The results indicated that production of leukotrienes, potent mediators of inflammatory reactions, are affected by the type of polyunsaturated fatty acids in the diet.

Animals↗

PubMed1987

Initiation of lipid peroxidation in biological systems.

The direct oxidation of PUFA by triplet oxygen is spin forbidden. The data reviewed indicate that lipid peroxidation is initiated by nonenzymatic and enzymatic reactions. One of the first steps in the initiation of lipid peroxidation in animal tissues is by the generation of a superoxide radical (see Figure 16), or its protonated molecule, the perhydroxyl radical. The latter could directly initiate PUFA peroxidation. Hydrogen peroxide which is produced by superoxide dismutation or by direct enzymatic production (amine oxidase, glucose oxidase, etc.) has a very crucial role in the initiation of lipid peroxidation. Hydrogen peroxide reduction by reduced transition metal generates hydroxyl radicals which oxidize every biological molecule. Hydrogen peroxide also activates myoglobin, hemoglobin, and other heme proteins to a compound containing iron at a higher oxidation state, Fe(IV) or Fe(V), which initiates lipid peroxidation even on membranes. Complexed iron could also be activated by O2- or by H2O2 to ferryl iron compound, which is supposed to initiate PUFA peroxidation. The presence of hydrogen peroxide, especially hydroperoxides, activates enzymes such as cyclooxygenase and lipoxygenase. These enzymes produce hydroperoxides and other physiological active compounds known as eicosanoids. Lipid peroxidation could also be initiated by other free radicals. The control of superoxide and perhydroxyl radical is done by SOD (a) (see Figure 16). Hydrogen peroxide is controlled in tissues by glutathione-peroxidase, which also affects the level of hydroperoxides (b). Hydrogen peroxide is decomposed also by catalase (b). Caeruloplasmin in extracellular fluids prevents the formation of free reduced iron ions which could decompose hydrogen peroxide to hydroxyl radical (c). Hydroxyl radical attacks on target lipid molecules could be prevented by hydroxyl radical scavengers, such as mannitol, glucose, and formate (d). Reduced compounds and antioxidants (ascorbic acid, alpha-tocopherol, polyphenols, etc.) (e) prevent initiation of lipid peroxidation by activated heme proteins, ferryl ion, and cyclo- and lipoxygenase. In addition, cyclooxygenase is inhibited by aspirin and nonsteroid drugs, such as indomethacin (f). The classical soybean lipoxygenase inhibitors are antioxidants, such as nordihydroguaiaretic acid (NDGA) and others, and the substrate analog 5,8,11,14 eicosatetraynoic acid (ETYA), which also inhibit cyclooxygenase (g). In food, lipoxygenase is inhibited by blanching. Initiation of lipid peroxidation was derived also by free radicals, such as NO2. or CCl3OO. This process could be controlled by antioxidants (e).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1986

Hydroperoxide metabolism in trout gill tissue: effect of glutathione on lipoxygenase products generated from arachidonic acid and docosahexaenoic acid.

The generation of oxygenated products from arachidonic acid and docosahexaenoic acid by the n-9 lipoxygenase of trout gill was monitored as a function of substrate concentration and added glutathione. In the absence of added glutathione up to 50% of the substrate consumed by the lipoxygenase was ultimately converted non-enzymatically to trihydroxy derivatives of the initial n-9 hydroperoxide enzyme product. The presence of added glutathione progressively increased conversion of the respective fatty acid hydroperoxides to the n-9 monohydroxy derivatives of arachidonic and docosahexaenoic acids while concomitantly decreasing the yield of trihydroxy derivatives, consistent with its role as a cosubstrate in the peroxidase reaction. The stability and net turnover of the lipoxygenase were also significantly improved by the addition of glutathione. The relative distribution of monohydroxy and trihydroxy products from either arachidonic acid or docosahexaenoic acid were similarly affected and equally sensitive to the glutathione concentration. These data suggest that in animals, the hydroperoxides of n-6 and n-3 polyunsaturated fatty acids generated by lipoxygenases are equally metabolized by the peroxide scavenging capabilities of the tissue.

Animals↗

PubMed1986

Lipoxygenase in trout gill tissue acting on arachidonic, eicosapentaenoic and docosahexaenoic acids.

Lipoxygenase activity was characterized in the gill tissue of fresh-water trout. Incubation of arachidonic acid with gill preparations yielded 12-hydroxyeicosatetraenoic acid as the major product, suggesting a 12-lipoxygenase. Eicosapentaenoic acid was similarly converted to the 12-hydroxyeicosapentaenoic acid. Both arachidonic acid and docosahexaenoic acid were converted with equal apparent velocities and affinities into single monohydroxy derivatives. Analyses of the hydroxy product of docosahexaenoic acid were consistent with 14-hydroxydocosahexaenoic acid. This enzyme activity was localized to the cytosolic fraction and displayed a broad pH optimum around pH 7. The enzyme was insensitive to the cyclooxygenase inhibitors indomethacin and aspirin but activity was strongly inhibited in the presence of the lipoxygenase inhibitors, SnCl2 (5 mM), esculetin (10 microM) and eicosatetraynoic acid (100 microM).

Animals↗

PubMedDate not supplied

Body composition of inactivated growth hormone (oMt1a-oGH) transgenic mice: generation of an obese phenotype.

The consequences of a 42 d exposure to elevated growth hormone (GH) on adipose tissue were assessed using the regulatable ovine metallothionein- ovine GH (oMt1a-oGH) transgene in male and female GH transgenic (TG) mice. Activation of transgene expression at 21 d of age followed by inactivation of transgene expression at 63 d of age (TG-on/off) increased individual white adipose tissue (WAT) depots and total body lipid stores in both males and females. WAT, expressed as a percentage of fasted body weight, did not differ in wildtype (WT) and continuously activated TG males and females up to 105 d of age, but was increased approximately 270% following inactivation of the transgene. Inguinal depot adipocytes were more numerous in both male and female TG +/- relative to WT or TG animals. The ensuring obesity was not accompanied by a decrease in thermogenic capacity of brown adipose tissue, as indexed by uncoupling protein quantity. GH transgene expression was accompanied by elevated insulin levels that were restored to WT levels upon cessation of transgene expression (p > 0.1). Early, transient exposure to elevated GH increased total body lipid by nearly threefold independent of gender; the increased lipid content was sustained and reflected WAT hypertrophy and hyperplasia. The oMt1a-oGH mouse provides a novel model of induced obesity in response to inactivation of a GH-transgene by the withdrawal of the transgene stimulus.

Adipocytes↗