PubMed Health⌕ Search

Biomedical subjects

J Leonardi

Publications and source records attributed to J Leonardi.

At least 19 recordsLinked to original sources
PubMed1999

Beneficial effects of gamma linolenic acid supplementation on nerve conduction velocity, Na+, K+ ATPase activity, and membrane fatty acid composition in sciatic nerve of diabetic rats.

Metabolic and vascular abnormalities are implicated in the pathogenesis of diabetic neuropathy. Two principal metabolic defects are altered lipid metabolism resulting from the impairment of delta-6-desaturase, which converts linoleic acid (LA) into gamma linolenic acid (GLA), and reduced nerve Na+, K+ ATPase activity. This reduction may be caused by a lack of incorporation of (n-6) fatty acids in membrane phospholipids. Because this ubiquitous enzyme maintains the membrane electrical potential and allows repolarization, disturbances in its activity can alter the process of nerve conduction velocity (NCV). We studied the effects of supplementation with GLA (260 mg per day) on NCV, fatty acid phospholipid composition, and Na+, K+ ATPase activity in streptozotocin-diabetic rats. Six groups of 10 rats were studied. Two groups served as controls supplemented with GLA or sunflower oil (GLA free). Two groups with different durations of diabetes were studied: 6 weeks with no supplementation and 12 weeks supplemented with sunflower oil. To test the ability of GLA to prevent or reverse the effects of diabetes, two groups of diabetic rats were supplemented with GLA, one group for 12 weeks and one group for 6 weeks, starting 6 weeks after diabetes induction. Diabetes resulted in a 25% decrease in NCV (P < 0.0001), a 45% decrease in Na+, K+ ATPase activity (P < 0.0001), and an abnormal phospholipid fatty acid composition. GLA restored NCV both in the prevention and reversal studies and partially restored Na+, K+ ATPase activity in the preventive treatment group (P < 0.0001). These effects were accompanied by a modification of phospholipid fatty acid composition in nerve membranes. Overall, the results suggest that membrane fatty acid composition plays a direct role in NCV and confirm the beneficial effect of GLA supplementation in diabetic neuropathy.

Journal Article↗
PubMed1997

Lack of endogenous estrogens effects on circulating adhesion molecule ICAM-1.

The serum vascular adhesion molecule ICAM-1 that is involved in atherogenesis was determined in fertile, postmenopausal and at term pregnant women. The aim of the study was to ascertain if the antiatherogenic estrogens action may involve this adhesion molecule. The serum ICAM-1 concentrations resulted similar in the three groups studied: 331 +/- 35 ng/ml, 333 +/- 28 ng/ml and 302 +/- 53 ng/ml in fertile, postmenopausal and pregnant subjects respectively, despite the very different estrogen plasma levels. These data, the first on the ICAM-1 serum levels in women with different natural hormonal milieu, demonstrate that estrogens antiatherogenic action is not involving ICAM-1.

Adult↗
PubMed1997

Anticardiolipin antibodies in pre-eclampsia and intrauterine growth retardation.

OBJECTIVE: To establish an association of anticardiolipin antibody (ACA) levels and pre-eclampsia or intrauterine growth retardation (IUGR). METHODS: Twenty-eight patients with pre-eclampsia, 28 with IUGR and 28 normotensive control group were matched for maternal age, race, weight, cigarette smoking, and parity. All had plasma anticardiolipin antibodies (GPL and MPL) detected by the modified enzyme-linked immuno-absorbent assay (ELISA) technique. RESULTS: No statistical significant difference in ACA values, both GPL and MPL, was found among the three groups studied Furthermore, none reached a value of ACA that could be considered clinically relevant (> 15). CONCLUSION: No association was found in anticardiolipin antibody levels between pre-eclamptic and IUGR versus the control group.

Adult↗
PubMed1993

Effect of anthracyclines on phospholipase A2 activity and prostaglandin E2 production in rat gastric mucosa.

The purpose of this study was to investigate in rats the effects of three anthracyclines, pirarubicin, doxorubicin and epirubicin on gastric prostaglandin E2 (PGE2) metabolism and phospholipase A2 (PLA2, EC 3.1.1.4) activity. The level of the membrane precursor, arachidonic acid, and the stability of the membrane were investigated by analysis of the composition of fatty acids. Enzymatic activities involved in the turnover of membrane phospholipids such as lysophospholipase (LPase, EC 3.1.1.5) and acyl-CoA lysophosphatidylcholine: acyltransferase (ACLAT, EC 2.3.1.23), and in the detoxification of lipid hydroperoxides, selenium-dependent glutathione-peroxidase (GSH-PX, EC 1.11.1.9) were measured after injection of the drugs for 4 consecutive days. Pirarubicin does not give rise to any changes in these activities but doxorubicin and epirubicin decreased PGE2 production and the activities of PLA2, LPase and ACLAT. GSH-PX activity was not changed by any of the drugs. The decrease in PLA2 activity does not seem to be related to variations in membrane lipid composition because the total phospholipids content was unchanged. The P/S (polyunsaturated/saturated) ratio increased in the doxorubicin group and decreased in the epirubicin group, and the unsaturation index was moderately modified. Arachidonic acid was increased only in the doxorubicin group. In vitro, PLA2 activity was not inhibited by the three drugs in the micromolar range. A marked inhibition was observed at 2.5 mM for pirarubicin and at 1.0 mM for doxorubicin and epirubicin. The Lineweaver-Burk representation showed that these inhibitions were of an uncompetitive type. Pirarubicin may therefore be considered to be an anthracycline without marked side-effects on gastric mucosa. However, the in vitro inhibition of PLA2 activity by anthracyclines does not fully explain the in vitro decrease in PLA2 specific activity observed after doxorubicin and epirubicin treatment, and in this context membrane structure modifications unconnected with the lipid composition can not be excluded. In vivo these phenomena may affect PGE2 synthesis, whose level was lower in the doxorubicin and epirubicin groups than in control group.

Animals↗
PubMed1992

Eicosapentaenoic and docosahexaenoic acids in cultured rat ventricular myocytes and hypoxia-induced alterations of phospholipase-A activity.

Hypoxia was reported to induce a decrease in phosphatidylcholine-hydrolyzing phospholipase activity (PC-PLA) in cultured rat cardiomyocytes. This work was intended to compare the influence of the presence of either eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA) in the phospholipids on the PC-PLA activity in normoxic and hypoxic conditions. The enrichment of the medium with EPA or DHA resulted in cell phospholipids containing about 2% or 22% DHA, respectively. These cells were then submitted for 3.5 h to either normoxia or hypoxia and the PC-PLA activities were assayed using [1-14C] dioleoyl-PC (pH 8.4 for PC-PLA2 and 4.9 for PC-PLA1). The results show that both enzymic activities are significantly higher in DHA-rich cardiomyocytes. Hypoxia induced a significant decrease in PC-PLA2 (about 25%) which was not statistically different between the two groups of cells. The hypoxia-induced decrease in PC-PLA1 was not found significant. In conclusion, the nature of the long chain n-3 polyunsaturated fatty acids in the phospholipids appears to contribute to the regulation of PC-PLA activity but not to influence its decrease during hypoxia.

Animals↗
PubMed1992

Heart and liver membrane phospholipid homeostasis during acute administration of various antitumoral drugs to the rat.

The purpose of this study was to investigate in the rat heart and liver the effects of an acute administration of three anthracyclines, doxorubicin, epirubicin and pirarubicin, and an anthracenedione, mitoxantrone, on the membrane peroxidative status, which was estimated by the composition of polyunsaturated fatty acids (PUFA), and on the activities of the enzymes involved in membrane repair processes and lipid hydroperoxide detoxification. Rats were injected for four consecutive days with the drugs or saline (control) and killed 24 hr after the last injection. All the drugs induced an increase in plasma thiobarbituric reactive substances and alpha-tocopherol concentrations, both expressed per milligram of plasma lipids. Plasma vitamin A was decreased by about a factor of two by all the drugs. The fatty acid profile in the heart lipids showed that the polyunsaturated species (20:4 n-6, 22:6 n-3) remained at the same or even higher levels after anthracycline treatment. This can be explained by the fact that the activities of the enzymes involved in either the recycling of membrane phospholipids, such as phospholipases A1 and A2 (EC 3.1.1.4 and EC 3.1.1.32), lysophospholipases (EC 3.1.1.5) and acylCoA:lysophosphatidylcholine acyltransferases (EC 2.3.1.23), or hydroperoxide detoxification, such as selenium-dependent glutathione peroxidase (GSH-PX, EC 1.11.1.9) and glutathione S-transferases (GSH-T, EC 2.1.5.18), were maintained at the same level of activity after the antitumoral treatment. In liver, membrane phospholipid levels of PUFA were maintained as well as the activities of phospholipid-metabolizing enzymes. GSH-PX activity was not affected whereas that of GSH-T was slightly lowered by the drugs. These results suggest that during acute antitumoral-induced lipid peroxidation of membranes, the multi-enzymatic complex of the immediate processes of repair and detoxification is fully operational, allowing the membrane to rapidly recover its functional status. The results are discussed in the context of the equivocal relationships between antitumoral-induced lipid peroxidation and cardiac disturbances.

1-Acylglycerophosphocholine O-Acyltransferase↗
PubMed1990

In vitro study of docosahexaenoic acid incorporation into phosphatidylcholine by enzymes of rat heart.

Several studies have shown that in animals fed fish oils, docosahexaenoic acid (DHA) is incorporated into cardiac phosphatidylcholines (PC) mainly at the expense of arachidonic acid. In this study we were interested in examining if the enzymatic system involved in the remodeling of membrane PC presented any selectivity for DHA in rat heart. The enzymes that were studied from sequential incubations carried out in parallel, were acyl-CoA synthetase (EC 6.2.1.3) and acyl-CoA:lysophosphatidylcholine acyltransferase (EC 2.3.1.23) (ACLAT). The heart preparations examined were homogenates of whole heart and of purified cultured rat ventricular myocytes. Results showed that ACLAT tended to preferentially incorporate into PC the polyunsaturated fatty acids of the n-6 series (+30%) rather than those of the n-3 series. DHA, however, inhibited the incorporation of arachidonic acid (AA) into PC by 50% at a molar ratio (DHA/AA) of 1.5. This phenomenon seems to be related to the competitive inhibition exerted by DHA on the thio-esterification of AA, a reaction catalyzed by acyl-CoA synthetase. This inhibitory effect appears to be dependent on the kinetic properties of the acyl-CoA synthetase toward DHA which, among the fatty acids examined, exhibited the lowest apparent Km and Vmax. It is suggested that the intracellular pool of DHA-CoA is the determinant species in altering the DHA composition of cardiac PC in animals given fish oils.

Animals↗
PubMed1990

Phospholipase A activity of cultured rat ventricular myocyte is affected by the nature of cellular polyunsaturated fatty acids.

Fatty acid composition of membrane phospholipids of cultured cardiomyocytes can be modified by the type of polyunsaturated fatty acids (n-3 or n-6 PUFA) constituting the culture medium. In this study, we investigated the effect of fatty acid modification on the activities of the key enzymes involved in the deacylation-reacylation cycle of membrane phospholipids. Results showed that cardiomyocytes grown in the presence of n-6 PUFA exhibited a higher specific alkaline phospholipase A (mainly A2) activity (+34%) and a moderately lower lysophospholipase activity (-17%) than when incubated with n-3 PUFA. AcylCoA:lysophosphatidylcholine acyltransferase, acid lysosomal phospholipase A1 and acylCoA synthetase activities were not significantly altered by changes in cellular PUFA composition. It was demonstrated that the differences between phospholipase A activities of the two types of cultured cells were linked neither to a differential leakage of enzyme nor to oxidative injury to the enzyme through blockage of essential sulfhydryl groups. One likely explanation is that the PUFA-induced changes in membrane composition alter membrane physical properties which, in turn, affect membrane-bound phospholipase A activity. Possible beneficial effects of the n-3 PUFA-induced changes on membrane stability are discussed.

Animals↗
PubMed1990

Lipid diet and enterocyte microsomal membrane fluidity in rats.

Rats were fed on diets more or less enriched with n-3 and n-6 unsaturated fatty acids, before removal of the small intestine. The global protein, cholesterol and phospholipid contents of enterocyte microsomes were measured. Fatty acids of the total lipid extracts were determined. Acyl coenzyme A: cholesterol acyl transferase (ACAT) was chosen as the enzyme whose activity reflects metabolic changes induced by lipid diets. Fluorescence measurements using diphenylhexatriene as the membrane probe were performed. As dietary fat may change the fatty acid composition of membranes, the order parameter S calculated from fluorescence measurements was studied with regard to dietary fatty acid composition. The S values, distributed over a large range, were not different between rat groups. They were positively correlated with the ratios of cholesterol and proteins to phospholipids and the molar percentage of saturated fatty acids. ACAT activity was negatively correlated with S. Variations in S values among rats, whatever the diet, could in part be attributed to individual factors.

Animals↗
PubMed1989

Effects of fish oil, corn oil and lard diets on lipid peroxidation status and glutathione peroxidase activities in rat heart.

In this study, we investigated the effect of various types of fats on heart lipid peroxidation status and on blood lipid parameters. Rats were fed either a low-fat diet (2.2% lard plus 2.2% corn oil), a corn oil diet (17%), a salmon oil diet (12.5%) supplemented with 4.5% corn oil, or a lard diet (15%) supplemented with 2% corn oil. All diets were supplemented with 1% cholesterol. Rats were fed for eight weeks. When compared with the low-fat diet, the salmon oil-diet intake resulted in a lower blood cholesterol, triglyceride and phospholipid concentrations (-50, -56 and -30%, respectively). Corn oil only tended to lower blood lipids; this decrease was significant for triglycerides only (-40%). The hypocholesterolemic effect of salmon oil diet is even more pronounced, if blood cholesterol values are compared with those of rats fed the lard diet. Heart lipid composition was not affected by dietary manipulations. Fatty acid composition of cardiac phosphatidylcholines and phosphatidylethanolamines, however, were altered by high-fat diets. In phosphatidylcholine, salmon oil induced a twelvefold decrease in the n-6/n-3 ratio and a 26% increase in the unsaturation index. For phosphatidylethanolamine, the n-6/n-3 ratio decreased 7.7-fold and the unsaturation index increased by 13%. A 50% decrease of the n-6/n-3 ratio was observed in animals fed the lard diet. Ultramicroscopic examination of ventricles revealed that those of the salmon oil group significantly accumulated lipofuscin-like or ceroid material, whereas this accumulation was barely detectable in hearts of the other groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Compared effect of n-3 and n-6 dietary fatty acids on rat intestinal acyl-CoA:cholesterol acyltransferase activity.

Polyunsaturated fatty acids are known to lower plasma cholesterol concentrations. We studied their effect on intestinal acyl-CoA:cholesterol acyltransferase (ACAT) activity in rats fed either salmon oil or corn oil (17% fat) with or without 1% cholesterol. After an 8-week feeding period we confirmed the hypolipidemic effect of salmon oil and we established its ability to stimulate ACAT activity in rats fed low-cholesterol diets. The most striking effect of 1% dietary cholesterol on ACAT activity was obtained in the control group (34% enhancement), whereas cholesterol supplementation had no effect on ACAT activity in the salmon oil group. The results enable us to suggest that n-3 fatty acids have an effect per se on ACAT activity; the regulation of enzyme activity by dietary cholesterol probably involves independent processes.

Animals↗
PubMed1988

Activities of some enzymes of phospholipid metabolism in cultured rat ventricular myocytes in normoxic and hypoxic conditions.

Phospholipid catabolism is thought to be one of the critical events in membrane injury during heart ischemia. In this work, the enzymes involved in phospholipid metabolism were studied in purified cultured ventricular myocytes in normoxic and hypoxic conditions. Purified ventricular myocytes exhibited an alkaline phospholipase A activity which had sn-2 specificity and which was calcium dependent, and an acid phospholipase A activity with sn-1 specificity. These cells also exhibited lysophospholipase and acyl-CoA/lysophosphatidylcholine acyltransferase activities. Oxygen deprivation of the myocardial cells for 4 h resulted in a sharp reduction of both phospholipase A2 and A1 activities. The activities of the other lipolytic enzymes were unaffected by hypoxia. Although hypoxia resulted in a marked increase of lactate dehydrogenase leakage in the bathing fluid, no additional release of the lipolytic enzymes and mitochondrial enzyme was observed. However, we noted an important alkaline phospholipase A2 leakage during normoxia. It is suggested that ventricular myocytes, under hypoxia, tend to prevent phospholipid degradation by reducing their phospholipase A activities.

1-Acylglycerophosphocholine O-Acyltransferase↗
PubMed1987

Effect of dietary lipids on the lipid composition and phospholipid deacylating enzyme activities of rat heart.

Rats were fed lard-enriched (17%) or corn oil-enriched (17%) diets and were compared with rats fed a low fat (4.5%) diet. Cardiac protein, DNA, phospholipid (PL) and fatty acid (FA) compositions were analyzed. Neutral phospholipase A, lysophospholipase and creatine kinase activities in the membrane and cytosolic compartments were also investigated. No significant modification of cardiac protein, DNA nor PL was observed among the three groups. Some alterations appeared in the FA composition. A lard-enriched diet induced a significant increase of 22:5n-3 and 22:6n-3 in heart phosphatidylcholine (PC) and phosphatidylethanolamine (PE), whereas a linoleic acid-rich diet induced a specific increase of 22:4n-6 and 22:5n-6 in these two major PL. Compared to rats fed the low fat diet, membrane-associated phospholipase A activity, measured by endogenous hydrolysis of membrane PC and PE, showed a significant increase (+45%) for both PL in rats fed corn oil. However, the activity of membrane-associated phospholipases, measured with exogenous [1-14C]dioleoyl PC, was not different among the three groups of rats. Cytoplasmic activity was decreased in rats fed corn oil, and lysophospholipase and creatine phosphate kinase activities were not significantly affected by diet. FA modification of the long chain n-6 FA induced by corn oil may be responsible for the observed increase in phospholipase activity. Physiological implications are suggested in terms of membrane degradation and prostaglandin production.

Animals↗
PubMed1986

Partial characterization of rat heart cytosolic phospholipase A1 and demonstration of essential sulfhydryl groups.

The cytosol of rat heart has been previously shown to contain phospholipase A activity in substantial amounts. This paper describes the isolation and partial purification of rat heart cytosolic phospholipase A. After homogenization of rat heart followed by centrifugation to remove membraneous material, the phospholipase A activity was isolated by ammonium sulfate precipitation and further purified by gel permeation chromatography with Sephadex G-100 in the presence of 5 mM taurodeoxycholate. Two peaks were isolated: a minor peak at the void volume and major peak corresponding to a molecular weight of 45,000. The molecular weight observed in HPLC gel permeation chromatography experiments was also Mr 45,000 and was not significantly affected by the nature of the detergent used. Phospholipase A was purified 77-fold over the crude cytosol. Further purification could not be attained due to lability of the phospholipase A activity. The enzyme is a phospholipase of the A1 type which does not require Ca2+ and lacks lipase or transacylase activity. It is unusual for the phospholipases A described to date, since it is inhibited by thiol reagents and is protected by beta-mercaptoethanol, suggesting the presence of essential sulfhydryl residues.

Animals↗
PubMed1983

Adsorption of pancreatic (pro)phospholipase A2 to various physiological substrates.

The adsorption of pancreatic phospholipase was studied in vitro in the presence of egg yolk lipoprotein emulsion, Intralipid emulsion, and milk fat globules. When the emulsions are incubated with bile salts, the latter dissociate a considerable fraction of the phospholipids initially associated with the emulsions, leading to the coexistence of an emulsified phase and a phase of mixed micelles. After the addition of pancreatic phospholipase A2, gel filtration shows that the enzyme was more than 90% bound to mixed micelles, regardless of the type of emulsion used. Comparable results were obtained by replacing the bile salts with human gallbladder bile. In parallel, pancreatic zymogen was never found to be bound to any of the lipid structures present (emulsion or mixed micelles). When the catalytic site of pancreatic phospholipase A2 was blocked with 4-bromophenacylbromide, there was no fixation on mixed micelles. Fixation was restored by the presence of lysolecithins and fatty acids in the incubation mixtures. The partial transformation of all emulsified substrates to mixed micelles by bile salts in vivo would thus lead to optimum activity of pancreatic phospholipase A2.

Adsorption↗
PubMed1980

Pancreatic phospholipase A2 hydrolysis of phosphatidylcholines in various physicochemical states.

Bile salts-phosphatidylcholines-cholesterol mixed micelles, native bile, egg yolk and intralipid emulsions were used as pancreatic phospholipase A2 (EC 3.1.1.4) substrates. The enzyme activity depends on the bile salt/phosphatidylcholine molar ratio. The enzyme had a low specific activity on bile phosphatidylcholines, because of the existence in native bile of a high bile salt/phosphatidylcholine molar ratio generating unfavorable conditions of hydrolysis, as demonstrated with mixed micelles. However when the bile salt/phosphatidylcholine molar ratio from bile was decreased to 2 : 1, enzyme activity increases up to an optimum. This optimal activity was about one third that observed when the substrate was mixed micelles. Under these optimal conditions a simultaneous hydrolysis of intralipid and bile phosphatidylcholine mixture shows comparable initial hydrolysis rates. During an extended incubation, however, nearly all intralipid phosphatidylcholines and only half the bile phosphatidylcholines were hydrolyzed by pancreatic phospholipase A2. Bile salts mixture or native bile desorb a portion of the phosphatidylcholines from the intralipid emulsion in optimally hydrolysable bile salts phosphatidylcholines mixed micelles. These micelles bind about 85% of the enzyme indicating that hydrolysis occurs primarily in the micellar phase. These results are discussed in terms of fat lipolysis in vivo.

Animals↗