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Partial conservation of the sn-2 position of dietary triglycerides in fasting plasma lipids in humans.

The authors investigated the effect of the position of fatty acids within dietary triglycerides on the composition of plasma lipids. Sixty volunteers consumed two diets of equal fatty acid compositions for 3 weeks each. In the palm oil diet 82% of palmitic acid was attached to the sn-1 and sn-3, and 18% to the sn-2 position of glycerol. In the diet rich in a palm oil analogue, Betapol, these figures were 35% and 65% respectively. Oleic and linoleic acid in palm oil were mainly in the sn-2 position, and in Betapol mainly in the sn-1 and sn-3 position. The proportion of palmitic acid in the 2 position of fasting plasma triglycerides was 10·2 g 100 g-1 on palm oil and 12·3 on Betapol; that of oleic acid was 46·dot 9 vs. 43·6 (P < 0·0001). Overall oleic acid content of triglycerides was 1·4 g 100 g-1 lower on Betapol than on palm oil (P = 0·002). Betapol increased palmitic and palmitoleic acid in cholesteryl esters by 1 g 100 g-1 (P < 0·0001) at the expense of oleic and linoleic acid. The positional configuration of dietary fatty acids has small but consistent effects on lipid metabolism that persist beyond fat absorption and chylomicron clearance.

Adult↗

Fluorine-19 nuclear magnetic resonance studies of lipid fatty acyl chain order and dynamics in Acholeplasma laidlawii B membranes. A physical, biochemical, and biological evaluation of monofluoropalmitic acids as membrane probes.

Fluorine-19 nuclear magnetic resonance spectroscopy offers a number of unique advantages for studies of lipid fatty acyl chain order and dynamics in model and biological membranes. However, the germinal difluoromethylene fatty acids commonly employed as 19F membrane probes appear to appreciably perturb the organization of model membranes and biomembranes. We have thus synthesized a series of specifically labeled monofluoropalmitic acids and evaluated these as suitable membrane probes. Differential scanning calorimetric studies of aqueous dispersions of several bis-(monofluoropalmitoyl)phosphatidylcholines reveal that a fluorine substitution near the carbonyl group of palmitic acid has only a modest effect on the thermotropic phase behavior of these model membranes and that substitutions in the center or toward the methyl terminus are relatively nonperturbing. Moreover, all bis(monofluoropalmitoyl)phosphatidylcholines tested exhibit nearly ideal mixing in all proportions with dipalmitoylphosphatidylcholine. The thermotropic phase behavior of membranes of the simple, cell-wall-less prokaryote Acholeplasma laidlawii B is also not detectably altered by the presence of appreciable amounts of biosynthetically incorporated monofluoropalmitic acid. We also find that the biosynthetic incorporation of even large amounts of monofluoropalmitic acids into the membrane lipids of A. laidlawii B has no effect upon the growth and survival of this organism. The presence of exogenous monofluoropalmitic acids in the growth medium does not alter the polar head group composition or lipid/protein ratio of the A. laidlawii B membrane. In addition, all monofluoropalmitic acids tested are biosynthetically incorporated as well as palmitic acid itself and distribute relatively evenly between the various membrane glyco- and phospholipids.(ABSTRACT TRUNCATED AT 250 WORDS)

Acholeplasma laidlawii↗

Measurement of palmitate availability in serum samples: method and utility.

Palmitic acid shows a very low and unknown solubility at neutral pH. Binding equilibria of palmitate to human serum albumin accordingly cannot be investigated by measuring free and bound ligand concentrations as in conventional binding studies. It is feasible, on the other hand, to describe the binding equilibria in relative terms, by measuring the concentration, p, of reserve albumin, previously defined as the concentration of a purified standard albumin preparation which in buffered solution binds a trace amount of palmitate as tight as it is bound in the sample. Palmitate availability is calculated as C/p, when C is the concentration of bound palmitate. The general binding equation is modified to contain the availability beside relative binding constants, Li = Ki/K1,St, where K1,St is the first stoichiometric binding constant for palmitate to the standard albumin preparation. Availabilities and relative binding constants can replace free concentrations and usual binding constants in considerations of biochemical transport and enzymatic mechanisms. A method is described for measuring the concentration of reserve albumin for binding of palmitate, based upon determination of dialytic exchange rates for palmitate among identical equilibrium samples. A technique for reproducibly adding radiolabelled palmitate to the samples is given.

Adult↗

Retinoylation of HL-60 proteins. Comparison to labeling by palmitic and myristic acids.

Recent studies suggest that a retinoic acid (RA) nuclear receptor or a retinoylated nuclear protein may be involved in the action of RA. We showed previously (Takahashi, N., and Breitman, T. R. (1989) J. Biol. Chem. 264, 5159-5163) that retinoylation involves the formation of a thioester bond and occurs on protein in newly formed cells and in pre-existing cells. In this study, we saw at least 14 retinoylated proteins in HL-60 cells. Greater than 90% of the retinoylation was associated with the nuclear protein described previously. This protein, partially purified from isolated nuclei, bound to DNA-cellulose and was eluted with NaCl. Retinoylation occurred in HL-60 cells exposed to cycloheximide. Thus, retinoylation resembled palmitoylation, both in the covalent bond and the exchangeable reaction involving preformed protein. These similarities prompted us to compare retinoylation with two other fatty acylations in growing HL-60 cells. We found that the major retinoylated protein was labeled by either radioactive palmitic acid or myristic acid. The extent of [3H]palmitic acid labeling of this protein was not reduced by growth in the presence of RA. The extent of retinoylation of this protein was not reduced by growth in the presence of increasing concentrations of palmitic acid. These results raise the possibility that the same protein is a substrate for retinoylation, palmitoylation, and myristoylation.

Cell Line↗

One-year effects of increasingly fat-restricted, carbohydrate-enriched diets on lipoprotein levels in free-living subjects.

Restriction of all dietary fat is a popular strategy for restricting saturated fat intake to lower LDL cholesterol. Some authorities advise the restriction of fat intake to the extreme of less than 10% of daily energy on the assumption that more fat restriction is better. The two studies described herein address questions relating to whether increasing fat restriction produces proportionally increasing benefit on cardiovascular risk factors in hyperlipidemic subjects. The first study is the Dietary Alternatives Study (DAS). The DAS was conducted in 531 male Boeing employees over a 2-year period. Subjects were defined as hypercholesterolemic (HC) or combined hyperlipidemic (CHL) based on age-specific 75th percentiles for plasma LDL-C and triglyceride levels. Hypothesis test analyses were performed at 1 year. HC subjects were randomized to diets taught to attain fat intakes of 30, 26, 22, and 18% (Diets levels 1-4, respectively). CHL subjects (slightly fewer in number) were randomized to Diets 1-3. After 1 year, subjects' total fat intakes were 27, 26, 25, and 22% of energy (en%), resulting in saturated fat intakes of 8, 7, 7, and 6%, respectively. In HC subjects the greatest LDL-C decrease was with Diet 2 (mean of 13.4%) and in CHL subjects with Diet 1 (7.0%). Surprisingly, plasma triglyceride concentrations rose in HC subjects 20% and 40% above baseline on Diets 3 and 4, respectively, with reciprocal reductions in HDL cholesterol of 2.5% and 3%, respectively. Furthermore, apo B reductions were attenuated below Diet 2 in HC subjects and Diet 1 in CHL subjects, and no further reductions were seen in plasma glucose and insulin concentrations, blood pressure, or body weight. Measurements of plasma total fatty acid composition showed a slight increase in plasma palmitate, whereas stearate decreased slightly, supporting the idea that de novo synthesis of palmitic acid was increased in the chronic high-carbohydrate feeding condition. The second study asked if the most effective diet in HC subjects, Diet 2, has an equivalent effect in women and men. To answer this question, men and women Boeing employees were taught the closely similar National Cholesterol Education Program (NCEP) Step II diet. After 6 and 12 months, equivalent reductions in LDL cholesterol were observed in women compared with men. HDL cholesterol levels in men were unchanged from baseline at 6 and 12 months, but were reduced 8% in HC women, with accompanying decreases of 18% in HDL2-cholesterol and 5% in apoprotein A-I (all P < 0.01). These data indicate that intakes of fat below about 25 en% and carbohydrate intake above approximately 60 en% yield no further LDL-C lowering in HC and CHL male subjects and can be counterproductive to triglyceride, HDL-C, and apo B levels. This lack of benefit appears to be explained by an enhanced endogenous synthesis of palmitic acid, which negates the benefit of further saturated fat restriction. The HDL-C decrease in HC women may have a similar cause and points to an underlying male-female difference. Alternative dietary approaches to limit saturated fat intake deserve intensive study.

Adult↗

Lipid peroxidation in the killing of phagocytized pneumococci.

To directly examine the role of hydrogen peroxide in the killing of bacteria after ingestion by granulocytes, we have studied some of the events of phagocytosis of a mutant strain of pneumococci which is relatively deficient in peroxide production. The hydrogen peroxide-deficient pneumococci and the otherwise identical wild type were grown with [(14)C]arachidonic and [(3)H]palmitic acid labels to label their lipids with unsaturated and saturated fatty acids, respectively. They were then incubated with both normal and chronic granulomatous disease granulocytes. The rates of ingestion and bacterial killing and the stability of fatty acids in the cell-bacteria complex were followed. Radioactive carbon dioxide released from glucose was also independently followed to measure glucose oxidation. Ingestion was similar for all cell-bacteria combinations. Chronic granulomatous disease cells killed the peroxide-positive wild pneumococci much more effectively (20-fold) than the peroxide-deficient mutant. Normal cells killed both peroxide-positive and -negative strains effectively. A considerable loss of [(14)C]arachidonic acid ( approximately 40%) consistent with lipid peroxidation of this unsaturated fatty acid was observed in all normal cells and in chronic granulomatous disease cells with peroxide-positive pneumococci. However, no loss of [(14)C]arachidonic acid occurred in chronic granulomatous disease cells with the peroxide-deficient pneumococci. No loss tritiated palmitic acid occurred in any cell-bacteria combination. Glucose oxidation was impaired in the chronic granulomatous disease cells in comparison to normal cells at rest and was especially impaired in chronic granulomatous disease cells ingesting the peroxide-deficient mutant pneumococci. This defect was partially corrected after phagocytosis of the peroxide-positive strain. These data directly support the hypothesis that bacterial killing is partially dependent upon an intact peroxide-generating system in the leukocyte-bacteria complex. Moreover, they indicate that bacterial lipid peroxidation is associated with the generation of peroxide during phagocytosis. Finally, they suggest that such peroxidation may contribute to effective phagocytic bacterial killing.

Animals↗

Alteration of fatty acid profiles in different pulmonary surfactant phospholipids in acute respiratory distress syndrome and severe pneumonia.

Impairment of alveolar surfactant function has been documented in the acute respiratory distress syndrome (ARDS) and in severe pneumonia (PNEU); however, the underlying mechanisms are not completely understood. In the current report we present a detailed analysis of fatty acid (FA) profiles of different surfactant phospholipid (PL) classes isolated from bronchoalveolar lavage fluids (BALF) and large surfactant aggregates (LSA) from mechanically ventilated patients with ARDS (n = 8), ARDS associated with lung infection (ARDS + PNEU, n = 9), and PNEU (n = 22). Healthy volunteers served as control subjects (n = 8). PLs were isolated by thin-layer chromatography, and the FA profile of each PL class was assessed by gas chromatography. In addition, the minimal surface tension (gamma min) of untreated LSA and of LSA after supplementation with additional dipalmitoylated phosphatidylcholine (DPPC) was analyzed (pulsating bubble surfactometer). As compared with control LSA, the percentage of palmitic acid in phosphatidylcholine (PC) was significantly decreased in all patient groups (ARDS 63.0 +/- 2.0%, ARDS + PNEU 64.6 +/- 4.9%, PNEU 65.6 +/- 1.5%, control subjects 80.1 +/- 1.7%), whereas the relative amount of unsaturated species in PC increased significantly in all groups. Phosphatidylglycerol (PG) and phosphatidylinositol (PI) presented similar FA profiles in control subjects, but differed in the patients. The FA pattern of sphingomyelin (SPH) and phosphatidylethanolamine (PE) displayed only minor changes under conditions of respiratory failure. As compared with control subjects a highly significant increase of gamma min from near zero to approximately 16 mN/m was observed in all patients and was found to be inversely correlated to the percentage of palmitic acid in PC of LSA or BALF. Accordingly, values for gamma min were significantly improved upon secondary supplementation of LSA with DPPC up to control values. We conclude that marked changes in the FA composition of the predominant surfactant PL classes occur, both in ARDS triggered by nonpulmonary events and PNEU. The marked reduction of palmitic acid in the PC fraction may be related to changes in surfactant function under these conditions.

Adult↗

Fatty acid and carotenoid composition of Rhodotorula strains.

Lipid content and composition of fatty acids with 6-25 carbon atoms were studied on strains of the 13 pink or red yeast species belonging to the genus Rhodotorula. The total amount of lipid represented an average of 13% of the dry weight. The neutral and polar lipid fractions were analyzed separately. For all the strains studied, the major fatty acids in both fractions were oleic, linoleic and palmitic acids, which formed 80% of the total number of fatty acids. A notable amount of arachidonic acid, a precursor of eicosanoid hormones, was found in R. acheniorum, R. aurantiaca and R. bacarum. Depending on the strain, 1-10 carotenoid pigments were detected; beta-carotene was always the major carotenoid present.

Carotenoids↗

Changes in fatty acids composition during seed growth and physicochemical characteristics of oil extracted from four safflower cultivars.

Fatty acid contents at different stages of maturity and physicochemical characteristics of oil extracted from mature seeds of four safflower cultivars (S208, S400, S541, and S303) were studied. Results indicated that for all cultivars both saturated and unsaturated fatty acid contents fluctuated with seed growth and development. Palmitic acid content decreased up to day 20 after which it started to increase for the S400, S541, and S303 cultivars, while for S208 it progressively decreased. Stearic acid fluctuated with seed growth and development. Oleic acid showed slight changes with seed growth and development, while linoleic acid decreased with seed growth and development for the S400, S541, and S303 cultivars while for S208 after day 30, it started to increase significantly. For all cultivars, glyceride contents varied among the cultivars with maximum values of 97.70, 2.80 and 0.20 for tri-, di- and monoglycerides, respectively. Physicochemical investigation of mature seed oils showed that the color, density, refractive index, free fatty acids, peroxide value, saponification value and unsaponifiable matter were similar for all cultivars while viscosity, iodine value, and acetone insoluble matter varied among the cultivars.

Chemical Phenomena↗

Characterization of a palmitoyl-acyl carrier protein thioesterase (FatB1) in cotton.

The relatively high level of palmitic acid (22 mol%) in cotton seeds may be due in part to a palmitoyl-acyl carrier protein (ACP) thioesterase (PATE), which prefers C16:0-ACP as its substrate. In embryo extracts, PATE activity was highest at the maximum rate of reserve accumulation (oil and protein), occurring about 30-35 d post anthesis. Thioesterase activity toward oleoyl-ACP was relatively similar at all developmental stages examined, but was considerably lower than the PATE activity. In developing seeds and in cotyledons and hypocotyls of seedlings, the PATE activity predominated. A cotton PATE cDNA clone isolated by screening a cDNA library with a heterologous Arabidopsis FatB1 probe has a 1.7-kb insert sequence with an open reading frame of 410 amino acids, lacking codons for the three N-terminal amino acids. The predicted amino acid sequence of the cotton PATE preprotein has a characteristic stromal-targeting domain and a 63% identity to the Arabidopsis long-chain acyl ACP-thioesterase FatB1 sequence. Alkaline blot hybridization of cotton genomic DNA with the Arabidopsis FatB1 probe suggested the presence of at least two FatB1 thioesterase genes in cotton. Relative cotton FatB1 transcript abundance was compared by RT-PCR and slot blot analysis in total RNA extracts from embryos, seedlings and leaves of mature plants. The cotton FatB1 mRNA apparently was expressed in all tissues but paralleled the profiles of PATE enzyme activity and seed oil accumulation in embryos.

Acyl Carrier Protein↗

Nutrient uptake into undifferentiated and differentiated HT-29 cells in culture.

HT-29 human colon carcinoma cells in culture have many characteristics of enterocytes, and these cells have been used by others to study intestinal drug and nutrient transport and metabolism. When grown in glucose-containing medium, HT-29 cells are largely undifferentiated (HT-29glu), but when grown in the absence of glucose but in the presence of galactose (HT-29gal), the population of cells is mostly differentiated. This study was undertaken with HT-29glu and HT-29gal cells to study the uptake of palmitic acid (16:0), linoleic acid (18:2), and cholesterol. The relationship between concentration and uptake of 16:0, 18:2, and cholesterol was linear in HT-29glu and HT-29gal cells, with the relative values of the slopes of this relationship being 18:2 > > 16:0 > > cholesterol. The rates of uptake of these lipids were at least three times higher in HT-29gal than in HT-29glu cells. In HT-29glu cells, the relative rates of uptake of the sugars at 32 mM were D-glucose = galactose > fructose > > alpha-methylglucose. Uptake of these sugars was much greater in HT-29gal than in HT-29glu cells. When 100 microM forskolin was added to the incubation medium for 7 days post-confluency, which stimulates the activity of adenylate cyclase and thereby increases the intracellular synthesis of cAMP, there was no effect on the uptake of the lipids or the sugars in either HT-29glu or HT-29gal cells. Thus, (i) differentiated HT-29gal cells transport larger amounts of lipids and sugars than do undifferentiated HT-29glu cells; (ii) forskolin has no effect on the uptake of lipids or sugars in these cells. This human cell culture system may be useful to study the in vitro transport of lipids, to establish the role of cell differentiation on these uptake processes, and to determine the potential role of selected intracellular signals.

Absorption↗

A synthetic surfactant based on a poly-Leu SP-C analog and phospholipids: effects on tidal volumes and lung gas volumes in ventilated immature newborn rabbits.

Available surfactants for treatment of respiratory distress syndrome in newborn infants are derived from animal lungs, which limits supply and poses a danger of propagating infectious material. Poly-Val-->poly-Leu analogs of surfactant protein (SP)-C can be synthesized in large quantities and exhibit surface activity similar to SP-C. Here, activity of synthetic surfactants containing a poly-Leu SP-C analog (SP-C33) was evaluated in ventilated premature newborn rabbits. Treatment with 2.5 ml/kg body wt of 2% (wt/wt) SP-C33 in 1,2-dipalmitoyl-sn-3-glycero phosphoryl choline (DPPC)-1-palmitoyl-2-oleoyl-sn-3-glycero phosphoryl choline (POPC)-1-palmitoyl-2-oleoyl-sn-3-glycero phosphoryl glycerol (POPG), 68:0:31, 68:11:20, or 68:16:15 (wt/wt/wt) suspended at 80 mg/ml gave tidal volumes (Vt) of 20-25 ml/kg body wt, with an insufflation pressure of 25 cmH2O and no positive end-expiratory pressure (PEEP), comparable to the Vt for animals treated with the porcine surfactant Curosurf. Nontreated littermates had a Vt of approximately 2 ml/kg body wt. The Vt for SP-C33 in DPPC-egg phosphatidylglycerol-palmitic acid [68:22:9 (wt/wt/wt)], DPPC-POPG-palmitic acid [68:22:9 (wt/wt/wt)], and DPPC-POPC-POPG [6:2:2 (wt/wt/wt)] was 15-20 ml/kg body wt. Histological examination of lungs from animals treated with SP-C33-based surfactants showed incomplete, usually patchy air expansion of alveolar spaces associated with only mild airway epithelial damage. Lung gas volume after 30 min of mechanical ventilation were more than threefold larger in animals treated with Curosurf than in those receiving SP-C33 in DPPC-POPC-POPG, 68:11:20. This difference could be largely counterbalanced by ventilation with PEEP (3-4 cmH2O). An artificial surfactant based on SP-C33 improves Vt in immature newborn animals ventilated with standardized peak pressure but requires PEEP to build up adequate lung gas volumes.

Amino Acid Sequence↗

Elucidation of carbon sources used for the biosynthesis of fatty acids and sterols in the trypanosomatid Leishmania mexicana.

Sterols are necessary for the growth of trypanosomatid protozoans; sterol biosynthesis is a potential target for the use and development of drugs to treat the diseases caused by these organisms. This study has used (14)C-labelled substrates to investigate the carbon sources utilized by promastigotes and amastigotes of Leishmania mexicana for the production of sterol [mainly ergosta-5,7,24(24(1))-trien-3beta-ol] and the fatty acid moieties of the triacylglycerol (TAG) and phospholipid (PL) of the organism. The isoprenoid precursor mevalonic acid (MVA) was incorporated into the sterols, and the sterol precursor squalene, by the promastigotes of L. mexicana. However, acetate (the precursor to MVA in most organisms) was a very poor substrate for sterol production but was readily incorporated into the fatty acids of TAG and PL. Other substrates (glucose, palmitic acid, alanine, serine and isoleucine), which are metabolized to acetyl-CoA, were also very poor precursors to sterol but were incorporated into TAG and PL and gave labelling patterns of the lipids similar to those of acetate. In contrast, the amino acid leucine was the only substrate to be incorporated efficiently into the squalene and sterol of L. mexicana promastigotes. Quantitative measurements revealed that at least 70-80% of the sterol synthesized by the promastigotes of L. mexicana is produced from carbon provided by leucine metabolism. Studies with the amastigote form of L. mexicana showed that in this case leucine was again the major sterol precursor, whereas acetate was utilized for fatty acid production.

Acetic Acid↗

[Studies on chemical constituents from the flowers of Carthamus tinctorius L].

OBJECTIVE: To investigate the chemical constituents from the flowers of Carthamus tinctorius. METHOD: The chemical constituents were isolated and repeatedly purified on silica gel column. They were identified and structurally elucidated by means of physio-chemical constants and spectral analysis. RESULT: Eight compounds were identifed as palmitic acid (I), 1-O-hexadecanolenin (II), trans-3-tridecene-5,7,9,11-tetrayne-1,2-diol (III), trans-trans-3,11-tridecadiene -5,7,9-triyne -1,2-diol (IV), coumaric acid (V), daucosterol (VI), apigenin (VIl) and kaempferol (VIII), respectively. CONCLUSION: Two compounds (II and III) were obtained from the genus of Carthamus for the first time.

Apigenin↗

Omega-3 fatty acids in baked freshwater fish from south of Brazil.

Lipid and fatty acid levels in the edible flesh of 17 baked freshwater fish from Brazil's southern region were determined. Analyses of fatty acids methyl esters were performed by gas chromatography. Palmitic acid (C16:0) was the predominant saturated fatty acid, accouting for 50-70% of total saturated acids. Linoleic acid (C18:2 omega 6), linolenic acid (C18:3 omega 3), and docosahexaenoic acid (C22:6 omega 3) were the predominant polyunsatured fatty acids (PUFA). The data revealed that species such as barbado, corvina, pintado, and truta were good sources of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), and that most freshwater fish examined were good sources of PUFA-omega 3.

Animals↗

Clinical evaluation of the 14C fat absorption test.

The absorption of (14)C-labelled glyceryl tripalmitate was tested in 62 patients by determining the maximum specific activity of CO(2) in the breath after an oral dose. A significant correlation with faecal fat excretion was found. The absorption of (14)C labelled palmitic acid was tested subsequently in 54 patients. In cases of pancreatic steatorrhoea, normal absorption of palmitic acid with impaired absorption of glyceryl tripalmitate was found, whereas in most patients with coeliac disease the absorption of both substances was depressed to the same extent.

Blind Loop Syndrome↗

Possible mechanism for formation and regulation of the palmitate-induced cyclosporin A-insensitive mitochondrial pore.

The mechanism of the palmitate-induced opening of the mitochondrial Ca2+-dependent cyclosporin A (CsA)-insensitive pore was studied, as well as the influence on this process of well-known modulators of the CsA-sensitive Ca2+-dependent pore. Palmitic acid, which can bind Ca2+ with high affinity, induced the cyclosporin A-insensitive swelling of mitochondria, whereas palmitoleic and 2-bromopalmitic acids, which have no such affinity for Ca2+, failed to induce the pore opening. The palmitate-induced Ca2+-dependent swelling of mitochondria was not affected by a well-known inhibitor of the CsA-sensitive pore (ADP) and an activator of this pore (inorganic phosphate, P(i)). However, this swelling was inhibited by physiological concentrations of ATP ([I]50 = 1.3 mM), but 100 microM ATP increased by 30% the rate of mitochondria swelling if Ca2+ had been added earlier. The effects of ATP (inhibition and activation) manifested themselves from different sides of the inner mitochondrial membrane. Mg2+ inhibited the palmitate-induced Ca2+-dependent swelling of mitochondria with [I]50 = 0.8 mM. It is concluded that palmitic acid induces the opening of the CsA-insensitive pore due to its ability for complexing with Ca2+. A possible mechanism of the pore formation and the influence of some modulators on this process are discussed.

Adenosine Triphosphate↗

Proton leak induced by reactive oxygen species produced during in vitro anoxia/reoxygenation in rat skeletal muscle mitochondria.

Superoxide anion generation and the impairment of oxidative phosphorylation yield were studied in rat skeletal muscle mitochondria submitted to anoxia/reoxygenation in vitro. Production of superoxide anion was detected after several cycles of anoxia/reoxygenation. Concomitantly, a decrease of state 3 respiration and phosphorylation yield (ADP/O) were observed. The latter resulted from a proton leak. The presence of palmitic acid during anoxia/reoxygenation cycles led to a dose-dependent inhibition of superoxide anion production together with a partial protection of the ADP/O ratio measured after anoxia/reoxygenation. The ADP/O decrease was shown to be due to a permeability transition pore-sustained proton leak, as it was suppressed by cyclosporine A. The permeability transition pore activation was induced during anoxia/reoxygenation by superoxide anion, as it was cancelled by the spin trap (POBN), which scavenges superoxide anion and by palmitic acid, which induces mitochondrial uncoupling. It can be proposed that the palmitic acid-induced proton leak cancels the production of superoxide anion by mitochondria during anoxia/reoxygenation and therefore prevents the occurrence of the superoxide anion-induced permeability transition pore-mediated proton leak after anoxia/reoxygenation.

Animals↗