PubMed Health⌕ Search

Biomedical subjects

G Nalbone

Publications and source records attributed to G Nalbone.

At least 37 records · Page 2Linked to original sources
PubMed1992

Effects of dietary n-6/n-3 ratios on lipid and prostaglandin E2 metabolism in rat gastric mucosa.

The effects of increased dietary n-3 polyunsaturated fatty acids on gastric mucosal lipid metabolism were studied in rats fed for 8 weeks with different combinations of fish and corn oils. Lipid composition, ex vivo prostaglandin E2 (PGE2) production and enzymatic activities involved in phospholipid metabolism and peroxisomal oxidative catabolism of fatty acids and PGE2 were examined. With dietary n-6/n-3 compositional ratios ranging between 75 and 3.3 it was observed that: (i) the arachidonic acid-to-eicosapentaenoic acid ratio (AA/EPA) fell from infinity to 3.1 and 5.1 in phosphatidylcholines (PC) and phosphatidylethanolamines (PE), respectively; (ii) ex vivo production of PGE2 was lowered by a factor of about 2; and (iii) gastric phospholipase A2 activity was enhanced by 32%. With dietary n-6/n-3 ratio lower than 3.3, stimulation of PGE2-CoA oxidase activity was observed whilst the PGE2 level remained constant. These data suggest that the fish oil-induced decrease in ex vivo PGE2 production is more closely related to a decrease in the membrane AA level than to an enhanced oxidative catabolism of PGE2.

1-Acylglycerophosphocholine O-Acyltransferase↗
PubMed1991

Subcellular localization of rat gastric phospholipase A2.

In the present study, we have performed experiments to gain some insight into the subcellular localization and biochemical properties of gastric mucosal phospholipase A2. After classical subcellular fractionation of whole glandular stomach mucosa, we found that gastric phospholipase A2 was essentially enriched in the 105,000 x g pellet that contains microsomes and plasma membranes. Except for the cytosol, all the subcellular fractions exhibited similar phospholipase A2 activity (i.e., optimum of pH, calcium dependence, apparent Km and positional specificity). The high-speed pellet was further characterized by ultracentrifugation on a sucrose gradient. Data showed that the sedimentation profile of phospholipase A2 was quite similar to those of plasma membrane markers and more specifically to an apical membrane marker. These results, taken together, showed that a gastric phospholipase A2 is distributed among the various subcellular fractions (as a result of cross-contamination) together with the membrane fraction on which it is associated. It is proposed that this fraction is the apical plasma membrane which would be the main site of phospholipase A2 action for arachidonic acid release. Lysophospholipase showed the same sedimentation profile as phospholipase A2, whereas acyl CoA-lysophosphatidylcholine: acyltransferase mainly sedimented with heavy microsomes. The substrate specificity of the enzyme was assessed by endogenous hydrolysis of gastric mucosal phospholipids. We were able to show that the enzyme acts at nearly the same rate on two major gastric membrane phospholipids, namely phosphatidylcholine and phosphatidylethanolamine.

Animals↗
PubMed1991

Modulation of lipid chylomicron-synthesizing enzymes in rats by the dietary (n-6):(n-3) fatty acid ratio.

The effect of diets with various (n-6):(n-3) polyunsaturated fatty acid (PUFA) ratios and a constant polyunsaturated: saturated fatty acid ratio on the basal activity of chylomicron lipid synthesizing enzymes was investigated in rat intestinal microsomes. Enzymes studied were: acyl-CoA:cholesterol acyltransferase (ACAT); acyl-CoA:lysophosphatidylcholine acyltransferase (MGAT) and acyl-CoA:1,2-diacylglycerol acyltransferase (DGAT). Results showed that after a 4-wk feeding period, ACAT, ACLAT and DGAT basal activities were significantly enhanced by the dietary fatty acids of the (n-3) series, whereas MGAT activity was not affected. When the highest (n-6):(n-3) ratio (51.0) was compared with the lowest (0.8), the increase attained was about 58, 76 and 73% for ACAT, ACLAT and DGAT, respectively. Fatty acid composition of microsomal lipids was drastically altered by the diets because (n-3) PUFA replaced the (n-6) series as the dietary (n-6):(n-3) ratio was lowered. These compositional changes could explain the observed modification in the membrane-bound enzyme activities. We suggest that (n-3) PUFA ingestion leads to an enhanced velocity of chylomicron synthesis in rats.

Acyltransferases↗
PubMed1990

Effect of increasing amounts of dietary fish oil on brain and liver fatty composition.

Increasing dietary fish oil in rat had the following effect on brain lipids: Arachidonic acid regularly decreased; eicosapentanenoic acid, normally nearly undetectable, was present; 22:5(n - 3), dramatically increased but remained below 1% of total fatty acids; cervonic acid was increased by 30% at high fish oil concentration. Saturated and monounsaturated fatty acids were not affected regardless of chain-length. In contrast, in the liver, nearly all fatty acids (saturated, monounsaturated and polyunsaturated) were affected by high dietary content of fish oil, but liver function was normal: serum vitamin A and E, glutathione peroxidase, alkaline phosphatase, transaminases were not affected. Serum total cholesterol, unesterified cholesterol and phosphatidylcholine were slightly affected. In contrast, triacylglycerols were dramatically reduced in proportion to the fish oil content of the diet.

Animals↗
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

Inverse modifications of heart and liver alpha-tocopherol status by various dietary n-6/n-3 polyunsaturated fatty acid ratios.

The effect of dietary n-6/n-3 fatty acid ratio on alpha-tocopherol homeostasis was investigated in rats. Animals were fed diets containing fat (17% w/w) in which the n-6/n-3 ratio varied from 50 to 0.8. This was achieved by combining corn oil, fish oil, and lard. The polyunsaturated to saturated ratio and total alpha-tocopherol remained constant in all diets. Results showed that enrichment of n-3 polyunsaturated fatty acids in the diet, even at a low amount (3.9% w/w), resulted in a dramatic reduction of blood alpha-tocopherol concentration, which, in fact, is the result of a decrease in plasma lipids, since the alpha-tocopherol to total lipids ratio was not significantly altered. The most striking effect observed was a considerable alpha-tocopherol enrichment (x 4) of the heart as its membranes became enriched with n-3 polyunsaturated fatty acids. This process appeared even with a low amount of fish oil (3.9% w/w) added to the diet. Accordingly, a strong positive correlation was found between heart alpha-tocopherol and docosahexaenoic acid (r = 0.86) or docosahexaenoic acid plus eicosapentaenoic acid levels (r = 0.84). Conversely, the liver alpha-tocopherol level dropped dramatically when n-3 polyunsaturated fatty acids were gradually added to the diet. It is concluded that fish oil intake dramatically alters the alpha-tocopherol homeostasis in rats.

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

Effects of dietary corn oil and salmon oil on the oxidation of fatty acids and prostaglandin E2 in rat gastric mucosa.

The investigations previously carried out by Grataroli and colleagues (1) to elucidate the relationships between dietary fatty acids, lipid composition, prostaglandin E2 production and phospholipase A2 activity in the rat gastric mucosa are, here, extended. In the present investigations, fatty acid and prostaglandin E2 catabolizing enzymes were assayed in gastric mucosa from rats fed either a low fat diet (corn oil: 4.4% w/w) (referred as control group), a corn oil-enriched diet (17%) or a salmon oil-enriched diet (12.5%) supplemented with corn oil (4.5%) (referred as groups of treated animals) for eight weeks. Peroxisomal fatty acyl-CoA beta-oxidation was induced in the treated animals whereas the activities of catalase and mitochondrial tyramine oxidase were increased and normal, respectively. Mitochondrial acyl-CoA dehydrogenations occurred at higher rates and carnitine acyltransferase activities were enhanced. In addition, the induction of peroxisomal but not mitochondrial prostaglandoyl-E2-CoA beta-oxidation could be demonstrated. Induction of peroxisomal oxidation of fatty acids and prostaglandins is suggested to contribute to the decrease of prostaglandin E2 production in the treated animals, especially those receiving the salmon oil diet, that the above mentioned authors originally reported.

Animals↗
PubMed1988

High dietary fish oil alters the brain polyunsaturated fatty acid composition.

Feeding adult rats a 17% corn-oil diet for 8 weeks did not change brain polyunsaturated fatty acids (PUFA) compared to rats fed 2.2% corn oil (with 2.2% lard added). When the corn-oil diet was supplemented with 14.5% cod liver oil or 12.5% salmon oil, the fatty acid composition of brain PUFA was significantly altered, even if alpha-tocopherol was added to the salmon-oil diet. Comparing salmon-oil- and cod-liver-oil-fed animals with corn-oil-fed animals, arachidonic acid 22:4(n-6) and 22:5(n-6) were reduced, and 20:5(n-3), 22:5(n-3) and 22:6(n-3) were increased. Liver fatty acids were also significantly altered. Thus, the brain is not protected against a large excess of very-long-chain n-3 PUFA, which increase n-3/n-6 ratio and could lead to abnormal function, and which might be difficult to reverse.

Animals↗
PubMed1988

Use of a silicic acid microcolumn to assay acyl-CoA: lysophosphatidylcholine acyltransferase.

A simple and rapid method for assaying acyl-CoA:lysophosphatidylcholine acyltransferase is described. This method is based on silicic acid microcolumn chromatography using labelled lysophosphatidylcholine (lysoPC) as substrate. The reaction was stopped by conventional Folch extraction. The chloroform extract (2 ml) was deposited on the silica gel and pushed through with air, and then elution was performed with methanol/water (50:50, v/v). Under these conditions, only the labelled phosphatidylcholine (PC) synthesized was retained on the gel, and this was then removed from the column and counted immediately. This method gave enzyme activities comparable to those obtained with the TLC method, and has proved to be reproducible. The new method, however, is both faster and safer than the classical TLC method.

1-Acylglycerophosphocholine O-Acyltransferase↗
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↗
PubMed1988

Effects of dietary corn oil and salmon oil on lipids and prostaglandin E2 in rat gastric mucosa.

Three groups of male rats were fed either a corn oil-enriched diet (17%, w/w), a salmon oil-enriched diet (12.5%) supplemented with corn oil (4.5%) or a low-fat diet (4.4%) for eight wk to investigate the possible relationships between dietary fatty acids and lipid composition, and prostaglandin E2 level and phospholipase A2 activity in the rat gastric mucosa. High-fat diets induced no important variation in total protein, phospholipid and cholesterol contents of gastric mucosa. Compared with a low-fat diet, corn oil produced a higher n-6/n-3 ratio in mucosal lipids, whereas this ratio was markedly lowered by a fish oil diet. In comparison with the low-fat diet, the production of prostaglandin E2(PGE2) in gastric mucosa of rats fed salmon oil was significantly decreased by a factor of 2.8. In the corn oil group, PGE2 production tended to decrease, but not significantly. In comparison with the low-fat diet, both specific and total gastric mucosal phospholipase A2 activities were increased (+ 18 and 23%, respectively) in the salmon oil group; they were unchanged in the corn oil group. It is suggested that the decrease of gastric PGE2 in rats fed fish oil is not provoked by a decrease in phospholipase A2 activity but may be the result of the substitution of arachidonic acid by n-3 PUFA or activation of PGE2 catabolism.

Animals↗
PubMed1988

Acyl-coenzyme A: cholesterol acyltransferase assay: silica gel column separation of reaction products.

Acyl-coenzyme A:cholesterol acyltransferase (ACAT) assays are usually performed by incubation of the enzyme with a labeled substrate followed by thin-layer chromatography separation and subsequent quantification of cholesteryl esters (CE) formed. Herein, a method is described for rapid separation of CE from other lipids, by elution from a silica gel column with a solvent mixture of petroleum ether/diethyl ether (98:2, v/v). Silica gel column chromatography is reliable and more rapid and safer than TLC. The best results were obtained when the reaction was stopped by Dole extraction followed by CE separation on a silica gel column. Assays for ACAT from rat intestinal microsomes showed that the specific activity values obtained using this method were reproducible and in good agreement with those obtained by conventional TLC method.

Animals↗
PubMed1988

Effect of dietary salmon oil feeding on rat heart lipid status.

For 2 mo rats were fed a salmon oil diet (12.5%, wt/wt) supplemented with 4.5% (wt/wt) corn oil, a corn oil diet (17%, wt/wt) or a low fat diet (4.4%, wt/wt). Cardiac lipids were analyzed and fatty acid composition of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) was determined. Ventricular biopsies were taken for ultramicroscopic examination. Serum cholesterol, triglyceride, phospholipid and vitamin E concentrations were significantly lower in rats fed salmon oil than in those fed the other two diets, whereas serum transaminases and vitamin A were not significantly affected. Cardiac protein, phospholipid, triglyceride and cholesterol concentrations were unaffected by diet. Cardiac phospholipid composition remained unchanged and no significant changes in lyso-PC or lyso-PE levels were observed. However, the salmon oil diet produced a markedly lower n-6/n-3 ratio in both PE and PC than in the other two diets. This was the result of replacement of n-6 polyunsaturated fatty acids (PUFA), primarily 20:4n-6 with n-3 PUFA, primarily 22:6n-3. The unsaturation index of PC and PE was higher with the salmon oil diet than with the other two diets. Ventricular biopsies of rats fed salmon oil showed mild lipid accumulation associated with some lipofuscin-like material. It is suggested that, in rat heart, fish oil led to a moderate accumulation of lipids, the composition of which may include long-chain monounsaturated fatty acids and a degradative form of peroxidized lipids.

Animals↗
PubMed1988

Subcellular distribution of lysophospholipase of rat intestinal mucosa.

We have studied the subcellular localization of rat intestinal lysophospholipase activity and some of the biochemical properties of this enzyme. After subcellular fractionation, an enriched activity was found in the high-speed pellet fraction containing the microsomes and the brush border membranes. Subsequently, these organelles were isolated. Using the classical calcium-precipitation method to isolate brush border membranes, we failed to demonstrate any significant recovery of lysophospholipase activity associated with this fraction. The microsomal fraction was further isolated after density gradient centrifugation, and most of the lysophospholipase activity was recovered with this fraction. Because further purification of the enzyme was unsuccessful, some of the biochemical properties of the enzyme were determined on the partially purified microsomal fraction. The optimum pH of the activity was centered at 7.0, and the enzyme did not require bivalent cations. By using double reciprocal plots, we determined the Kapp(m) to be 0.4 mM; the Vapp(max), 23 mumol.h-1.mg protein-1. The enzyme was strongly inhibited by detergents having a low critical micellar concentration and less inhibited by those having a higher critical micellar concentration.

Animals↗
PubMed1988

Effect of bile anionic polypeptidic fraction on the fate of cholesterol carried by liposomes in the rat.

[14C]Cholesterol associated with liposomes with or without anionic polypeptidic fraction was administered intravenously to the rat. The cholesterol originated from liposomes including anionic polypeptidic fraction is secreted in bile much later, is stored in liver in higher quantity, and is metabolized into bile salts in lesser quantity during the 4 hr of experimentation than the cholesterol issued from liposomes exempt of anionic polypeptidic fraction. From these results it can be postulated that the cholesterol associated with liposomes containing anionic polypeptidic fraction might be directed in a particular liver pathway.

Animals↗