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Intestinal absorption of Mn-mesoporphyrin in a small bowel sac system: effect of oleic acid.

RATIONALE AND OBJECTIVES: The authors investigated the effect of oleic acid (cis-9-octadecenoic acid) (OA), a lipidic carrier, on the intestinal absorption rate and T1 relaxation time of manganese (III) mesoporphyrin (Mn-mesoporphyrin), a prototype hepatobiliary contrast agent for magnetic resonance imaging. METHODS: Mn-mesoporphyrin was formulated with OA at various concentrations. Small bowel sacs were created in 36 rats and filled with complexed and free Mn-mesoporphyrin. Intestinal absorption of Mn-mesoporphyrin was measured with spectrophotometry at 364 nm. T1 relaxation times were measured in samples of Mn-mesoporphyrin solutions, bowel wall, liver, and bile. RESULTS: Absorption rates ranged from 4.2%/cm2/h to 13%/cm2/h. Absorption was greatest (13%/cm2/h) when a combination of 1 mmol/L Mn-mesoporphyrin and 26.5 mmol/L OA was used. The T1 of bile decreased from 2,480 to 248 msec (maximum decrease) in rats that received Mn-mesoporphyrin. CONCLUSION: Mn-mesoporphyrin is absorbed from the small bowel in both the lipid-associated and free form, resulting in substantial shortening of the T1 in bile.

Animals↗

Saccharomyces cerevisiae PIP2 mediating oleic acid induction and peroxisome proliferation is regulated by Adr1p and Pip2p-Oaf1p.

Saccharomyces cerevisiae genes involved in fatty acid degradation contain in their promoters oleate response elements (OREs) and type 1 upstream activation sequences (UAS1s) that bind Pip2p-Oaf1p and Adr1p, respectively. The promoter of the PIP2 gene was found to contain a potential UAS1 that consists of a tandem array of CYCCRR half-sites in an overlapping arrangement with a previously characterized ORE. Electrophoretic mobility shift analysis demonstrated that Adr1p bound to UAS1PIP2, and Northern analysis in combination with a lacZ reporter gene confirmed that Adr1p influenced the transcription of PIP2. Immunoprecipitation showed that, in adr1delta mutant cells grown on oleic acid, Pip2p was less abundant compared with the corresponding wild-type. In addition, the amount of Pip2p-Oaf1p that bound to a target ORE in vitro was reduced in mutant extracts compared with the wild-type. Transcription of the oleic acid-inducible genes SPS19 and CTA1, which rely on both Pip2p-Oaf1p and Adr1p for their regulation, was reduced in adr1delta mutant cells. However, by ectopically restoring levels of Pip2p in adr1delta cells grown on oleic acid medium, transcription of both genes increased 2-fold compared with the control. This partial suppression of the adr1delta mutant phenotype was additionally manifested by moderate utilization of oleic acid. Hence, both the expression as well as the action of the two transcription factors, Adr1p and Pip2p-Oaf1p, are interconnected, which allows for an elaborate control of fatty acid-inducible genes.

Blotting, Northern↗

Probing interfacial chemistry of single droplets with field-induced droplet ionization mass spectrometry: physical adsorption of polycyclic aromatic hydrocarbons and ozonolysis of oleic acid and related compounds.

The recently developed technique of field-induced droplet ionization (FIDI) is applied to study interfacial chemistry of a single droplet. In a new variation of the FIDI method, 1-2-mm-diameter droplets hang from a capillary and undergo heterogeneous reactions between solution-phase analytes and gas-phase species. Following a specified reaction time, the application of a high electric field induces FIDI in the droplet, generating fine jets of highly charged progeny droplets that are characterized by mass spectrometry. Sampling over a range of delay times following exposure of the droplet to gas-phase reactants, the spectra yield the temporal variation of reactant and product concentrations. We illustrate the technique with three examples: the adsorption of the polycyclic aromatic hydrocarbon naphthalene into a water-methanol droplet, the ozonolysis of oleic acid, and localization of the carbon-carbon double bond within a lysophosphatidic acid. Gas-phase naphthalene reacts with 80% methanol-20% water droplets containing 100 microM silver nitrate. Positive ion mass spectra show increasing concentrations of silver ion-naphthalene adducts as exposure times increase. To examine the ozonolysis of organic molecules, gas-phase ozone generated by a mercury pencil-style lamp reacts with either 10 microM oleic acid or 100 microM oleoyl-L-alpha-lysophosphatidic acid (LPA; 18:1). Negative ion spectra from the ozonolysis of oleic acid show azelaic acid and 9-oxononanoic acid as the principle reaction products. Ozonolysis products from LPA (18:1) unambiguously demonstrate the double bond position in the original phospholipid.

Journal Article↗

Modification of plasma and hepatic lipids of guinea pigs by feeding high oleic acid pork compared with regular pork.

Meat from such monogastric animals as swine can be modified to substitute monounsaturated fatty acids for saturated fatty acids. Because monounsaturated fatty acids have a beneficial effect on serum lipids as compared with saturated fatty acids, the objective of this study was to assess the effect of modified pork as compared with regular pork on serum and hepatic lipids. Guinea pigs were fed diets containing pork from control diet-fed hogs or from hogs fed a diet containing high oleic acid sunflower oil. The pork provided almost all of the fat in the diets at the level of 4 and 15 g/100 g diet, 10 or 34% energy. The high oleic pork muscle and fat contained 26 and 46% less palmitic and stearic acids (the primary saturated fatty acids), respectively, and 31 and 29% more oleic acid (the primary monounsaturated fatty acid) than the regular pork muscle and fat, respectively. Cholesterol concentration of diets ranged from 0.06 to 0.08% of the diet. Although total serum cholesterol, HDL cholesterol and triglyceride concentrations did not differ due to type of pork, results indicated that serum LDL cholesterol was lower (15%) and hepatic cholesterol was greater (15%) in the high oleic pork, 15% fat group as compared with the control pork 15% fat group. Also, serum LDL cholesterol concentration was higher in the groups fed 15% fat compared with those fed 4% fat. In this study pork modified to have more oleic acid and less saturated fatty acids had a positive effect on tissue lipids when fed to animals.

Animals↗

Fc gamma of IgG: a specific agent of destabilization of lipid bilayers containing oleic acid.

gamma-Immunoglobulins induce the fusion of oleic acid-containing liposomes into tubular structures. An F(ab')2 preparation does not exert the same influence unless it is several orders of magnitude more concentrated. The same is true for several proteins with an isoelectric point higher or lower than the IgG isoelectric point. The Fc of IgG seems thus to exert a specific destabilizing and fusogenic action on artificial lipid membranes. An hypothesis is presented concerning the mode of action of Fc gamma on lymphocyte membrane which is based on the facts mentioned above and on the existence of a phospholipasic activity of Fc gamma membrane receptor.

Fluorescence↗

Breathing patterns in lambs after oleic acid lung injury utilizing respiratory inductive plethysmography.

We measured breathing patterns utilizing a respiratory inductive plethysmograph (RIP) in seven healthy nonsedated lambs after an iv infusion of oleic acid (50 mg/kg) to induce acute pulmonary edema. Our single position graphic (SPG) calibration technique was employed for gain factor calculation. Accuracy was validated by the simultaneous volume measurement of RIP and integrated pneumotachography (PNT). Of a total 840 validation breaths, 467 (56%) were within 5% of PNT, 734 (87%) were within 10%, and 834 (99.9%) were within 20%. In each study baseline physiologic and breathing pattern data were collected and also at 15, 30, 60, 90, 150, and 210 min postoleic acid infusion. Validation of RIP accuracy before each data collection revealed 29% required new gain factor calculation. Recalibration was done within 5 min. Excluding respiratory frequency, which remained at 30% above baseline, variables were not significantly different than baseline measurements at the 210-min interval. Results suggest that calibration of RIP using our SPG technique is a time-efficient method and that RIP can accurately measure breathing patterns, providing an additional tool for assessment of experimental lung injury in lambs.

Animals↗

Oleic acid increases cell surface expression and activity of CD11b on human neutrophils.

Traumatic bone injury frequently results in the release of marrow-derived fatty material into the circulation. This may lead to the syndrome of fat embolism, associated with the generation of free fatty acids, the sequestration of neutrophils in the lungs, and the subsequent development of acute respiratory distress. Neutrophil accumulation in tissues requires their adherence to vascular endothelial cells and involves the beta2 integrin, CD11b/CD18 (Mac-1). We now report that the exposure of isolated human neutrophils to oleic acid causes a rapid increase in the cell surface expression and affinity state of CD11b, particularly under acidic conditions that are typical of inflammatory sites. Oleic acid also triggers neutrophil aggregation and neutrophil adherence to both fibrinogen-coated surfaces and confluent cultures of HUVEC. These processes are blocked by CD11b-specific inhibitors, including neutrophil-inhibitory factor and mAbs to CD11b. These observations may help explain the etiology of so-called fat embolism wherein trauma-induced release of fatty material causes pulmonary neutrophil accumulation and the development of acute respiratory distress.

Cells, Cultured↗

[Comparison in genetically hyperlipoproteinemic and normal rats of the captation and incorporation of isotopic oleic acid and glycerol at high concentration by the perfused liver].

Perfusions of isolated livers from genetically hyperlipoproteinemic Zucker fa/fa and normolipemic Zucker Fa/- rats are performed with loads of ]9,10-3H2] oleic acid and [1-14C] glycerol. The hepatic acylglycerols anabolism from these precursors is higher in the fa/fa rat than in the control Fa/- rats. Synthesis by esterification (of oleic acid) is more increased than de novo synthesis (from glycerol). The increase in lipid anabolism is due to an augmentation of the hepatic cellular mass, but this anabolism is not regulated in the same way than in the normal rat.

Animals↗

Superoxide production by rat neutrophils in the oleic acid model of lung injury.

The purpose of this study was to investigate the superoxide anion (O2-)-generating capacity of neutrophils isolated from rats at various stages of oleic acid(OA)-induced lung injury. Neutrophils were collected from blood, bronchoalveolar lavage (BAL), and peritoneal cavity (glycogen induced) after OA administration. Control neutrophils were collected from the blood of normal animals as a representative of nonprimed cells that produce low levels of O2-. A second control was the glycogen-elicited peritoneal neutrophil of normal animals which represented primed cells that produce enhanced levels of O2-. The ability of the neutrophils to produce O2- was evaluated by using both myristate acetate and opsonized zymosan as stimulants. Neutrophils isolated from blood and BAL from OA-injured lungs produced low levels of O2- and resembled closely the circulating, nonprimed neutrophil. Myeloperoxidase levels were measured in plasma and BAL and were found to be elevated in BAL of OA-injured animals. The inability of neutrophils to produce high levels of O2- and the elevation of myeloperoxidase suggest that neutrophils present in the lung may have degranulated in response to prior activation and are therefore incapable of further superoxide production.

Animals↗

Molten globule of bovine alpha-lactalbumin at neutral pH induced by heat, trifluoroethanol, and oleic acid: a comparative analysis by circular dichroism spectroscopy and limited proteolysis.

The calcium-depleted form of alpha-lactalbumin (alpha-LA) at neutral pH can be induced to adopt a partly folded state or molten globule upon moderate heating, by dissolving the protein in aqueous TFE or by adding oleic acid. This last folding variant of the protein, named HAMLET, can induce apoptosis in tumor cells. The aim of the present work was to unravel from circular dichroism (CD) measurements and proteolysis experiments structural features of the molten globule of apo-alpha-LA at neutral pH. CD spectra revealed that the molten globule of apo-alpha-LA can be obtained upon mild heating at 45 degrees C, as well as at room temperature in the presence of 15% TFE or by adding to the protein solution 7.5 equivalents of oleic acid. Under these various conditions the far- and near-UV CD spectra of apo-alpha-LA are essentially identical to those of the most studied molten globule of alpha-LA at pH 2.0 (A-state). Proteolysis of the 123-residue chain of apo-alpha-LA by proteinase K at 4 degrees C occurs slowly as an all-or-none process leading to small peptides only. At 37 degrees C, proteinase K preferentially cleaves apo-alpha-LA at peptide bonds Ser34-Gly35, Gln39-Ala40, Gln43-Asn44, Phe53-Gln54, and Asn56-Asn57. All these peptide bonds are located at level of the beta-subdomain of the protein (chain region 34-57). Similar sites of preferential cleavage have been observed with the TFE- and oleic acid-induced molten globule of apo-alpha-LA. A protein species given by the N-terminal fragment 1-34 linked via the four disulfide bridges to the C-terminal fragment 54-123 or 57-123 can be isolated from the proteolytic mixture. The results of this study indicate that the same molten globule state of apo-alpha-LA can be obtained at neutral pH under mildly denaturing conditions, as indicated by using a classical spectroscopic technique such as CD and a simple biochemical approach as limited proteolysis. We conclude that the molten globule of alpha-LA maintains a native-like tertiary fold characterized by a rather well-structured alpha-domain and a disordered chain region encompassing the beta-subdomain 34-57 of the protein.

Animals↗

Acute cardiopulmonary effects of nitroglycerin in canine oleic acid pulmonary edema.

In a canine model of acute respiratory failure, the authors investigated acute cardiopulmonary effects of nitroglycerin (TNG) and compared the results with those obtained after phlebotomy. Oleic acid increased intrapulmonary shunt (Qs/Qt) from 7.4 to 31% (P less than 0.001) and decreased (P less than 0.01) cardiac output (CO). In the presence of assumed low-pressure pulmonary edema, TNG was infused to decrease mean blood pressure (BP) by 40%; this was associated with a 26% decrease (P less than 0.05) in CO. Qs/Qt increased from 31 to 42% (P less than 0.01). There was a slight increase (P less than 0.01) in pulmonary vascular resistance (PVR) with TNG, and mean pulmonary artery pressure (PAP) decreased (P less than 0.05). In contrast, when CO was decreased by a similar amount with phlebotomy, mean Qs/Qt did not significantly change. There were similar changes in PVR and PAP and mixed venous O2 tension with TNG and phlebotomy. Accordingly, current results rule out increased flow, increased PVO2, and mechanical alterations in pulmonary vascular pressures as contributory to the increase in Qs/Qt with TNG. Alternatively, the increase in Qs/Qt with TNG may be explained by a direct pharmacologic decrease in pulmonary hypoxic vasoconstriction and/or by nonspecific pharmacologic effects.

Animals↗

Oleic acid supplementation reduces oxidant-mediated dysfunction of cultured porcine pulmonary artery endothelial cells.

We have previously shown that supplementing cultured porcine pulmonary artery endothelial cells (PAEC) with exogenous oleic acid (18:1 omega 9) alters the fatty acid composition of the cells and reduces oxidant-mediated cytotoxicity. Because the mechanisms by which lipid alterations modulate oxidant susceptibility have not been defined, the ability of 18:1 to reduce hydrogen peroxide (H2O2)-mediated PAEC dysfunction was evaluated. PAEC monolayers on polycarbonate filters were incubated for 3 h in maintenance medium supplemented with either 0.1 mM 18.1 in ethanol vehicle (ETOH) or with an equivalent volume of vehicle alone. Twenty-four hours later monolayers were treated for 30 min with 50 or 100 microM H2O2 in Hanks' balanced salt solution (HBSS) or with HBSS alone (nonoxidant control). As a functional index of PAEC monolayer integrity, the permeability of monolayers to albumin was then measured for 3 h. Treatment with 100 microM H2O2 caused cytotoxicity and progressive increases in PAEC monolayer permeability that were attenuated by 18:1 supplementation, whereas 50 microM H2O2 caused only a transient increase in permeability without cytotoxicity. Supplementation with 18:1 also attenuated H2O2-induced reductions in PAEC adenosine triphosphate (ATP) content and disruption of PAEC microfilament architecture. The ATP content of PAEC monolayers was reversibly reduced in the absence of oxidant stress by incubation with glucose-depleted medium containing deoxyglucose and antimycin A. Metabolic inhibitor-induced ATP depletion increased monolayer permeability and altered cytoskeletal architecture, alterations that resolved during recovery of PAEC ATP content. These results demonstrate that ATP depletion plays a critical role in barrier dysfunction and suggests that the ability of 18:1 to reduce oxidant-mediated PAEC dysfunction and injury may relate directly to its ability to preserve PAEC ATP content.

Actin Cytoskeleton↗

Association of changes in lysophosphatidylcholine metabolism and in microsomal membrane lipid composition to the pulmonary injury induced by oleic acid.

Alterations in the lipid composition of lung microsomal membranes occur in oleic acid-induced respiratory distress. The marked decrease in the phosphatidylcholine/lysophosphatidylcholine molar ratio could be related with an altered metabolism of lysophosphatidylcholine in these membranes. Results revealed that the activity of phospholipase A increased whereas that of acyl-CoA:lysophosphatidylcholine acyltransferase decreased. Microsomal lysophospholipase activity remained unchanged. On the other hand, the microsomal enzyme system involved in the de novo synthesis of diacylglycerol was impaired, and cholinephosphotransferase activity was lowered. These changes in the activity of some membrane-bound enzymes were not caused by changes in the membrane lipid fluidity since lipid structural order parameter (SDPH) did not change and neither did the major factors on which the fluidity depends. The possible significance of microsomal lipid alterations in the pathogenesis of respiratory distress induced by oleic acid is discussed.

1-Acylglycerophosphocholine O-Acyltransferase↗

Effect of different pressure levels on the dynamics of lung collapse and recruitment in oleic-acid-induced lung injury.

The effects of different inspiratory and expiratory airway pressures (Paw) on the dynamics of lung collapse and recruitment were studied in 14 anesthetized, mechanically ventilated, pigs with oleic-acid-induced lung injury. Repetitive CT scans of the same slice were obtained every 0.8 s during different inspiration and expiration hold procedures. The mean lung density and amount of atelectasis were measured in each scan. Inspiration to a Paw of 15 cm H2O above PEEP resulted in recruitment of collapsed lung tissue, mainly within 1.4 s. During expiration lung density increased rapidly and at an almost even rate within the first 1.4 s, whereas a rapid increase of atelectasis occurred after an initial delay period of 0.6 s with PEEP = 10 or 15 cm H2O. PEEP of 20 or 25 cm H2O almost prevented lung collapse during expiration. Thus, in order to avoid cyclic alveolar collapse during mechanical ventilation in oleic-acid-induced lung injury, a PEEP level >= 20 cm H2O or an expiration time <= 0.6 s is required. Long inspiratory time intervals, as used in inverse ratio ventilation, seem to be of minor importance for the recruitment of collapsed lung tissue in this experimental model.

Analysis of Variance↗

Reduction of serum cholesterol and low-density lipoprotein cholesterol levels in a juvenile population after isocaloric substitution of whole milk with a milk preparation (skimmed milk enriched with oleic acid).

OBJECTIVE: To study the effects of serum lipid levels by isocaloric substitution of whole milk intake in a group of children with a milk preparation (fat-free milk enriched with oleic acid containing a small amount of cholesterol) that is frequently consumed in our community. DESIGN: A crossover clinical trial was carried out with random selection. There were 88 children of both sexes, ranging in age from 3 to 9 years, selected at a Primary Care Center. The children were divided into two homogeneous groups. The first group consumed whole milk for the first 7-month study period, whereas the second group drank the milk substitute. At the end of the first test period, serum lipid levels were measured. Then the type of diet was reversed: Group 1 children consumed the milk substitute whereas Group 2 children drank whole milk. The rest of their intake did not vary throughout the study. At the end of the second 7-month period we measured serum lipid levels again. The levels of serum lipid evaluated were total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, triglycerides, Apoprotein A1, Apoprotein B, and lipoprotein (a). For the statistical analysis the nonpaired and paired t tests were used. RESULTS: The mean level of cholesterol after taking whole milk was 4.53 mmol/L (175.26 mg/dl), and the mean level after taking the milk preparation was 4.2 mmol/L (162.65 mg/dl), which indicates a 7.2% decrease. The mean level of low-density lipoprotein cholesterol after whole milk intake was 2.73 mmol/L (106.1 mg/dl), whereas after consuming the milk preparation it was 2.47 mmol/L (96.1 mg/dl), which indicates a decrease of 9.5%. Triglycerides were reduced from 0.83 mmol/L (73.53 mg/dl) after whole milk to 0.72 mmol/L (63.79 mg/dl) after the milk substitute, which suggests a 13.25% decrease. High-density lipoprotein cholesterol, apoprotein A1, apoprotein B, and lipoprotein (a) did not undergo any significant change. CONCLUSIONS: To reduce serum levels of total cholesterol and low-density lipoprotein cholesterol, without reducing caloric intake, it may be beneficial to substitute the whole milk customarily consumed by children with a milk preparation of fat-free milk enriched with oleic acid.

Animals↗

Comparison between intra- and extracellular surfactant in respiratory distress induced by oleic acid.

The present study compares the phospholipid distribution and protein content in bronchoalveolar lavage, purified extracellular surfactant and lamellar bodies isolated from rabbits killed at intervals of 2.5, 12 and 24 h after oleic acid administration. The data suggest that the alteration of pulmonary surfactant could be partially due to the type II cell response to the injury.

Animals↗

Ventilation by high-frequency oscillation in rabbits with oleic acid lung disease.

The feasibility and efficiency of ventilation by high-frequency oscillation (HFO) were examined in animals with diffuse hemorrhagic lung disease. Twenty-four hours after injection with 0.12 ml/kg oleic acid, 11 spontaneously breathing rabbits had a mean (+/- SD) arterial O2 partial pressure (PaO2) of 65 +/- 16 Torr and arterial CO2 partial pressure (PaCO2) of 38 +/- 7 Torr [inspired fractional O2 concentration (FIO2) of 0.21]. Following paralysis animals were ventilated using a high-frequency oscillator for periods of 20 min followed by three successive hyperinflations to prevent atelectasis. Maintaining a constant mean airway pressure (MAP) of 6 cmH2O and fresh gas flow (FGF) of 2 1/min (FIO2 = 0.21), all combinations of frequency (5, 10, 20, and 30 Hz) and stroke volume (Vs) 2.6, 5.0, and 8.9 ml) were tested. At each frequency, an increase in Vs tended to lower mean PaCO2. At each Vs, CO2 elimination appeared maximal at 20 Hz, an effect attributable to decreasing effective Vs with increasing frequency. With constant Vs, MAP, and frequency, increasing FGF from 1 to 2 or 61/min decreased mean PaCO2 (P less than 0.05). With constant Vs, frequency, and FGF, increases in MAP from 2 to 10 cmH2O increased mean PaO2 (P less than 0.05). HFO, coupled with periodic hyperinflation, supports satisfactory gas exchange in rabbits with oleic acid lung injury. The efficiency of gas exchange is improved by independent increases in Vs, FGF, MAP, or frequency.

Animals↗