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Effect of dietary fat and cholesterol on uptake of oleic acid and triolein by everted sacs of bovine small intestine.

The influence of dietary fats on in vitro lipid absorption by bovine intestine was studied in 14 calves. Holstein bull calves were fed for 16 wk five liquid diets containing skim milk plus either 3.5% milk fat, 3.5% tallow, 3.5% tallow and .2% cholesterol, 7.0% tallow, or 7.0% tallow and .2% cholesterol. Uptake of oleic acid or triolein by everted jejunal or ileal sacs was measured after incubation for 30 min at 37 degrees C in pH 7.4 micellar solutions containing tritium-labeled oleic acid or tritium-labeled triolein. Lipids were extracted from homogenates of sacs and separated into lipid classes by thin layer chromatography. Equal amounts of oleic acid were taken up by jejunal or ileal sacs. Triolein uptake was less than oleic acid uptake, but uptakes by jejunal or ileal sacs did not differ. Oleic acid incorporation into triglyceride was three to four times greater in intestinal sacs from milk fat-fed calves than in sacs from calves fed either 3.5 or 7% tallow. Oleic acid incorporation in intestinal sacs from calves fed cholesterol as well as tallow was equal to that in milk fat-fed calves. Intestinal uptake and metabolism of oleic acid proceeded faster when calves were fed milk fat than when fed tallow. The lower intestinal incorporation with tallow increased if cholesterol was fed, suggesting that cholesterol either stimulated absorption or increased esterification.

Animals↗

Increased intestinal absorption of oleic acid with aging in the rat.

Changes in nutrient absorption could be responsible for some of the disorders associated with aging. Oleic acid is the most common dietary fatty acid. Therefore, we investigated its absorption by the small intestine of aging rats in vivo. We used a single pass intestinal perfusion technique to study absorption in animals between 6 and 138 weeks of age. Rats less than 70 weeks of age absorbed 14,000 to 15,000 nmol of oleic acid per 100 cm/hr. In contrast, rats 94 weeks of age or older, absorbed 22,000 to 23,000 nmol of oleic acid per 100cm/hr. The 61% increase in oleic acid absorption as the rats aged correlated with a decrease in the resistance of the unstirred water layer from 0.8 to 0.5 min/cm3/100cm. Concomitantly, the surface area of the unstirred water layer increased from 97.1 to 141.2 cm2/100cm. The changes in unstirred water layer resistance and surface area explain the observed increase in absorption of oleic acid with aging. The increased surface area and decreased resistance of the unstirred water layer are both conducive to increased intestinal absorption of this lipid nutrient as the animals aged.

Aging↗

Accelerating effect of 12-keto oleic acid on lipid peroxide and fluorescent productions in mouse liver homogenate.

12-Keto oleic acid is generally considered to be a secondary degradation product of peroxides produced by lipid peroxidation. The acid was added to a mouse liver homogenate or to a bovine serum albumin and its influences on lipid peroxidation and fluorescent production were investigated and compared with other acids. Just as with vitamin E deficiency, 12-keto oleic acid was shown to increase lipid peroxide formation and fluorescence productiod directly and indirectly. The increase of lipid peroxide formation was caused directly through the increase of the free radical production, and indirectly through change of the biomembrane structure. The fluorescence production increase was caused directly by reaction of the 12-keto oleic acid itself, and indirectly by acceleration of the lipid peroxidation.

Animals↗

Neuronal differentiation is triggered by oleic acid synthesized and released by astrocytes.

Unlike in the adult brain, the newborn brain specifically takes up serum albumin during the postnatal period, coinciding with the stage of maximal brain development. Here we report that albumin stimulates oleic acid synthesis by astrocytes from the main metabolic substrates available during brain development. Oleic acid released by astrocytes is used by neurons for the synthesis of phospholipids and is specifically incorporated into growth cones. Oleic acid promotes axonal growth, neuronal clustering, and expression of the axonal growth-associated protein-43, GAP-43; all these observations indicating neuronal differentiation. The effect of oleic acid on GAP-43 synthesis is brought about by the activation of protein kinase C, since it was prevented by inhibitors of this kinase, such as H-7, polymyxin or sphingosine. The expression of GAP-43 was significantly increased in neurons co-cultured with astrocytes by the presence of albumin indicating that neuronal differentiation takes place in the presence of oleic acid synthesized and released by astrocytes in situ. In conclusion, during brain development the presence of albumin could play an important role by triggering the synthesis and release of oleic acid by astrocytes, which induces neuronal differentiation.

Albumins↗

Carp oil or oleic acid, but not linoleic acid or linolenic acid, inhibits tumor growth and metastasis in Lewis lung carcinoma-bearing mice.

I examined the effects of carp oil, oleic acid, linoleic acid and linolenic acid on tumor growth and metastasis to the liver in mice implanted intrasplenically with highly metastatic Lewis lung carcinoma (LLC) tumors. Carp oil (0.1 or 0.2 mL per mouse) significantly reduced tumor growth and metastasis to the liver. Carp oil at 100 or 1000 mg/L inhibited the DNA synthesis in LLC cells, the capillary-like tube formation of human dermal microvascular endothelial cells (HMVEC) at 1000 mg/L and the adherence of LLC cells to HMVEC at 10 to 1000 mg/L (in vitro). Carp oil (0.2 mL per mouse) inhibited the angiogenesis induced by Matrigel supplemented with vascular endothelial growth factor (VEGF) and heparin (in vivo). Antitumor and antimetastatic actions of carp oil might be partly attributable to the inhibition of DNA synthesis in LLC cells and angiogenesis through the inhibition of the adherence of LLC cells to the microvascular endothelium. Oleic acid (0.1 or 0.2 mL per mouse) significantly inhibited the metastasis to the liver, but it had no effect on the primary solid-tumor growth. Oleic acid inhibited the angiogenesis in both in vitro and in vivo models. Oleic acid at 1000 micromol/L inhibited the DNA synthesis in LLC cells but did not affect the DNA synthesis in HMVEC. These inhibitory actions of oleic acid may be attributable to the inhibition of angiogenesis induced by the tumor. Linoleic acid and linolenic acid had no effect on tumor growth or metastasis to the liver.

Animals↗

Biosynthesis of a defensive insect alkaloid: epilachnene from oleic acid and serine.

The biosynthesis of the azamacrolide epilachnene by the coccinellid beetle Epilachna varivestis has been studied with 2H-labeled oleic acid, 2H-labeled L-serine, and 13C,15N-labeled L-serine. The incorporation of these precursors into epilachnene defines the origin of the alkaloid's entire carbon/nitrogen skeleton. GC/MS and GC/IR studies of alkaloid produced by Epilachna fed with deuteriated oleic acid show that oleic acid loses four carbon atoms from its carboxyl end during the biosynthesis. Other details, including the mechanism of carbon-nitrogen bond formation between the fatty acid and amino acid moieties, remain to be established.

Alkaloids↗

Aerosol products, mechanisms, and kinetics of heterogeneous reactions of ozone with oleic acid in pure and mixed particles.

Reactions of O3 with pure and mixed oleic acid particles and bulk solutions were investigated using a thermal desorption particle beam mass spectrometer. The results provide information on the effect of particle matrix on reaction products, mechanisms, and kinetics. The major aerosol products are alpha-acyloxyalkyl hydroperoxides, secondary ozonides, alpha-alkoxyalkyl hydroperoxides, and oxocarboxylic acids formed primarily through reactions of Criegee intermediates with products or with particle matrix compounds. For example, it is estimated that for the reaction of pure oleic acid particles with O3 the aerosol products consist of approximately 68% organic peroxides, 28% 9-oxononanoic acid, and 4% azelaic acid. Although the reaction rate of pure oleic acid particles corresponds to an atmospheric lifetime of minutes, reactions in liquid/solid particle matrices can be orders of magnitude slower. The peroxide products are relatively stable when exposed to matrices typical of atmospheric particles, indicating that the lifetimes of these compounds in the atmosphere may be long enough to allow for long-range transport.

Aerosols↗

Hydration of oleic acid by a Selenomonas strain from the ovine rumen.

A Selenomonas sp., isolated from the ovine rumen, was characterized with regard to its ability to hydrate oleic acid to 10-hydroxystearic acid. Hydration occurred only in stationary phase in a medium containing 0.1%, 0.5% (w/v) galactose or 0.5% (w/v) glucose, but not in a medium containing 1% galactose. Growth under a hydrogen headspace did not result in the production of stearic acid, the biohydrogenated product of oleic acid. Linoleic and linolenic acids (0.1% v/v) were not hydrated. It is concluded that the growing bacterium is unlikely to contribute to oleic acid hydration in the rumen.

Culture Media↗

Separation and quantitation of mono-, di-, and triglycerides and free oleic acid using thin-layer chromatography with flame-ionization detection.

alpha-Monoolein was prepared from glycidol and oleic acid by the regioselective opening of glycidol in the presence of an anionic resin. During the reaction, the lipochemical synthesis medium becomes enriched in monoolein, the effective emulsifying agent. This mixture can be analyzed by thin-layer chromatography coupled with flame-ionization detection (FID). The products and reagents do not need to be derivatized. Diglyceride and triglyceride by-products affecting the selectivity of the reaction also could be detected using this technique. Cholesterol was used as an internal standard. The factors influencing the separation, including the hydrogen flow rate, scan speed, and the composition of the developing solvent, were investigated. The degree of separation is highly sensitive to the hexane/diethyl ether ratio of the developing solvent. Good separation of triglyceride, oleic acid, the two diglycerides, cholesterol, alpha-monoolein, and glycidol was obtained with the mixture hexane/diethyl ether/formic acid (65:35:0.04, by vol). Detector response, detection limits, and rod-to-rod variations also were examined. A range of rod loads giving a straightforward relationship between FID response and amount of compound loaded (relative to oleic acid and alpha-monoolein) was defined. The accuracy of the quantification was illustrated by analysis of a mixture of oleic acid and alpha-monoolein standards of known composition.

Chromatography, Thin Layer↗

The effect of phosphatidylcholine and lysophosphatidylcholine on the absorption and mucosal metabolism of oleic acid and cholesterol in vitro.

The absorption and mucosal metabolism of [14C]oleic acid and [3H]cholesterol were studied using everted sacs of rat jejunum in an in vitro incubation system. The labeled compounds were present in the incubation mixture either singly or together as mixed micelles with bile salt and monoacylglycerol and in the presence or absence of phosphatidylcholine or lysophosphatidylcholine. The presence of cholesterol or phosphatidylcholine markedly suppressed oleic acid absorption. We suggest that both compounds interacted with the micelles causing changes in micellar mass, charge or configuration leading to possible interference with access of the fatty acid to the cell membrane. Lysophosphatidylcholine enhanced oleic acid absorption and stimulated incorporation of the fatty into mucosal triacylglycerol. When the incubation temperature was lowered to suppress metabolism lysophosphatidylcholine had no effect. The results suggest that the increased absorption occurring at the higher temperature was secondary to enhanced glycerol acylation. Lysophosphatidylcholine had only a minimal effect on cholesterol absorption and no effect on cholesterol acylation. Evidence is presented showing that lysophosphatidylcholine is itself well absorbed and variously metabolized. We conclude that phosphatidylcholine and lysophosphatidylcholine have quite divergent effects on lipid absorption but the full elucidation of their mechanisms of action must await further study.

Animals↗

Effect of oleic acid on arginine-induced glucagon secretion by the isolated perfused rat pancreas.

The isolated perfused rat pancreas was used to investigate the effect of oleic acid on glucagon secretion in response to 10 mmol/l arginine. In the absence of oleic acid and at 2.5 mmol/l calcium, arginine induced a biphasic glucagon secretion. At lower extracellular calcium concentration (1.0 mmol/l), the second phase of glucagon release was reduced, the first phase being unchanged. In the presence of 1,500 mumol/l oleic acid, the glucagon response to arginine was also biphasic, but second phase release was markedly inhibited, the first phase glucagon release being unchanged. Such an effect was not obtained when oleic acid concentration in the medium was 750 mumol/l. These results demonstrate that high concentrations of oleic acid inhibit glucagon secretion in response to arginine from the isolated perfused rat pancreas and support the concept that circulating free fatty acid levels are involved in the control of glucagon secretion.

Animals↗

Embolic pneumopathy induced by oleic acid. A systematic morphologic study.

This paper presents a systematic study of acute and chronic pulmonary lesions resulting from a single intravenous injection of oleic acid and a new fibrosis lung model is proposed: pulmonary interstitial fibrosis is obtained by means of a number of oleic acid intravenous injections. Nineteen adult dogs received 0.045 g/kg or 0.09 g/kg of oleic acid. A systematic morphologic study was carried out after 1, 2, 3, 4, 6, 12, 24, and 48 hours and 1, 2 and 4 weeks. Eleven other adult dogs received weekly one injection of 0.09 g/kg of pure oleic acid over a period of 1 to 3 months. Examination of the lung was carried out by means of light and electron microscopy and morphometry. An early stage characterized by the formation of thrombosis and cellular necrosis was followed by a repair stage with the proliferation of Type 2 cells and fibrotic foci in the subpleural areas. Lipid staining with Sudan IV allowed the onset and disappearance of lipid-laden macrophages to be ascertained. The late stage showed pulmonary fibrosis. The extent of the lesions is related to the number of oleic acid injections. Since interstitial pulmonary fibrosis invariably appeared, and only 2 dogs out of 11 died, the model is satisfactory for pathologist and physiologist.

Animals↗

Pulmonary edema: a CT study of regional changes in lung density following oleic acid injury.

Transmission computed tomography (CT) was used to study the global and regional density changes in the dog lung associated with oleic acid induced lung injury. The same level of the lower thorax was scanned (5 s/scan) during suspended ventilation at functional residual capacity prior to and after oleic acid infusion (0.05 ml/kg) into the right atrium. The first signs of edema were usually seen within 15 to 30 min after oleic acid infusion and consisted of patchy areas of increased density primarily in the peripheral and dependent zones. Mean CT density (Hounsfield units + 1,000) of lung cross sections from five dogs was 198 +/- 9 (SEM) during base line; density significantly (p less than 0.05) increased to 243 +/- 14 30 min after infusion and reached an apparent plateau of 294 +/- 31 75 min after oleic acid infusion. Thermal-dye dilution measurements indicated that extravascular lung water increased by 3.8 ml/kg from base line to 75 min after oleic acid infusion.

Absorptiometry, Photon↗

Acetic, propionic, and oleic acid as the possible factors influencing the predominant residence of some species of Propionibacterium and coagulase-negative Staphylococcus on normal human skin.

The minimum inhibitory concentration (MIC) of acetic and propionic acid for resident bacteria on normal human skin, such as Propionibacterium acnes and Staphylococcus epidermidis, was 25 mg/mL or more at any pH tested (pH 5.5-6.8). While the MIC of these acids for most of the transient bacteria was markedly decreased by lowering the pH of the media and at pH 5.5, the mean pH value of the normal human skin, the MIC was 6.25 mg/mL or less. The MIC of oleic acid for some strains of Gram-positive transient bacteria of Streptococcus, Micrococcus, or Bacillus was 100 micrograms/mL or less at all pH's tested. Staphylococcus aureus was resistant to this acid at pH 6.8, but became as sensitive as Streptococcus when the pH was lowered. The growth of P. acnes, the most predominant resident bacterium, was enhanced markedly and reached a maximum level at 6.25 mg/mL of propionic acid, 12.5 mg/mL of acetic acid, and 50-100 micrograms/mL of oleic acid. On the basis of these results, we presumed that acetic, propionic, and oleic acids are factors influencing the predominant residence of some species of Propionibacterium and coagulase-negative Staphylococcus on normal human skin.

Acetates↗

Dietary oleic acid not used during brain development and in adult in rat, in contrast with sciatic nerve.

In order to determine exactly the effect on the nervous system of concentration of dietary oleic acid on the fatty acid composition of different part of the nervous system, triglycerides were synthesized using chemical and enzymological methods. The dose-effect was determined using an experimental protocol with seven groups of rats who received a diet in which the oleic acid level varied from 0 to 6000 mg per 100 g diet, but the other ingredients were identical (in particular the essential fatty acids, linoleic and alpha-linolenic acid). Rats were fed the diets from two weeks before mating, and their pups were sacrificed aged either 21 or 60 days. When the level of oleic acid in the diet was increased, the main modifications observed in 21-day-old deficient animals were as follows. (i). For 18:1(n-9), in liver, plateau was reached at about 4 g oleic acid per 100 g diet. Below this level, the higher the dose the greater the response. In whole brain, brain myelin, and nerve endings (but not sciatic nerve) the oleic acid level remained optimal and constant whatever the level of oleic acid in the diet. (ii). 16:1(n-7) concentration decreased in liver and in sciatic nerve, but not in nervous tissue. (iii). In 60-day-old animals, results were generally similar to those in 21-day-old animals.

Age Factors↗

[Effect of epigallocatechingallate on acute lung injury induced by oleic acid in mice].

AIM: To investigate the effect of epigallocatechingallate (EGCG) on acute lung injury induced by oleic acid in mice and the possible mechanism. METHODS: Acute lung injury was induced by oleic acid in mice. Light microscopy and electron microscopy were used to examine histological changes and lung index as well as wet to dry weight ratio was calculated. Serum TNF-a level was measured by enzyme linked immunosorbent assay (ELISA) and the phosphorylation of p38 MAPK was determined by Western blotting. RESULTS: Pretreatment of EGCG significantly alleviated oleic acid induced lung injury accompanied by reduction of lung index and wet to dry weight ratio, decreased of TNF-a level in serum and inhibition of phosphorylation of p38 MAPK. CONCLUSION: EGCG showed beneficial effect on acute lung injury induced by oleic acid in mice. The ultimate reduction of TNF-alpha in serum caused by inhibition of phosphorylated p38 MAPK is involved in the mechanism of action of EGCG.

Animals↗

Pulmonary vasoconstriction in oleic acid induced lung injury. A morphometric study.

Distribution and severity of active vasoconstriction of muscular pulmonary arteries were morphometrically assessed in anaesthetized, paralysed and mechanically ventilated pigs with respiratory distress, induced by oleic acid. Vasoconstriction was deduced from the medial thickness which was measured and expressed as a percentage of external diameter. Six pigs received oleic acid (0.12 +/- 0.07 ml/kg), dissolved 1:1 in 96% alcohol, in multiple injections of 0.1 ml. Six pigs were used as controls. After the oleic acid injections a stable hypoxaemia (PaO2 = 57 +/- 8 mmHg, at an inspiratory oxygen fraction of 0.6) and pulmonary hypertension (mean Ppa = 36 +/- 2 mmHg) were obtained for several hours. Electron microscopy revealed swelling of endothelial cells with signs of degeneration. Medial thickness was far greater in the oleic acid group than in the control group; overall mean values were 8.1 +/- 3.2 and 3.8 +/- 1.7% respectively (P < 0.001). Arteries with prominent vasoconstriction were lying in clusters. This pattern was the same in dependent and non-dependent regions. We concluded that in oleic acid induced respiratory distress active vasoconstriction of muscular pulmonary arteries is an important factor in the development of pulmonary hypertension. Besides vasoconstriction, endothelial swelling and intravascular clotting may contribute to the development of pulmonary hypertension.

Animals↗

Meclofenamate enhances blood oxygenation in acute oleic acid lung injury.

To assess the role of vasoactive prostanoids in acute lung injury, we studied 16 dogs after intravenous injection of oleic acid (OA; 0.08 ml/kg). Animals were ventilated with 100% O2 and zero end-expiratory pressure. Base-line hemodynamic and blood gas observations were obtained 90-120 min following OA. Observations were repeated 30 min after infusion of meclofenamate (2 mg/kg; n = 10), or after saline (n = 6). Resistance to pulmonary blood flow was assessed using the difference between pulmonary arterial diastolic and left atrial pressures (PDG). Ventilation-perfusion (VA/Q) distributions were derived with the multiple inert gas technique. Prior to infusion, there were no significant differences between the two groups. PDG was elevated mildly above normal levels, and shunt flow was the principal gas exchange disturbance. Saline induced no significant changes in hemodynamics or gas exchange. Meclofenamate enhanced PDG to a small, significant degree and effected a 32% reduction in shunt flow (P less than 0.01). Perfusion was redistributed to normal VA/Q units with little change in low VA/Q perfusion or in overall flow. Arterial PO2 rose from 75 +/- 36 to 184 +/- 143 Torr (P less than 0.05). At autopsy, there were no significant differences in wet to dry lung weights. Prostaglandin inhibition redistributes perfusion from shunt to normal VA/Q units, thereby improving arterial PO2, without altering lung water acutely.

Animals↗