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[Binding of spin-labeled palmitic acid by bovine serum albumin].

Investigation of the binding of spin-labeled palmitic acid and its esters with bovine serum albumin is presented. Both probes are shown to bind strictly to strongly and loosely binding centers of protein. The quantitative study allows to determine the binding constant and number of protein binding sites. It is found that the total binding constant for esterified derivative is several times as much as that of palmitic acid.

Electron Spin Resonance Spectroscopy↗

Palmitic acid in HDL is associated to low apo A-I fractional catabolic rates in vivo.

BACKGROUND: HDL becomes enriched with non-esterified fatty acids (NEFAs) in some pathologies, such as nephrotic syndrome, as well as after aerobic exercise. However, little is known about the impact of NEFAs on HDL metabolism. We investigated the effects of one NEFA, the palmitic acid, on HDL structure and catabolism. METHODS: HDL enrichment with palmitic acid (HDLPal) was performed by fusing phosphatidyl choline small unilamellar vesicles containing the NEFA with human HDL isolated from a pool of 5 normolipidemic plasma. HDL enriched only with phosphatidyl choline (HDLPhl) and native HDL (HDLCtrl) were included as controls. RESULTS: As expected, HDLPal surface charge density was higher than HDLPhl and HDLCtrl (2014.4+/-164.8 vs. 1682.7+/-149.5 and 1758.2+/-124.3-esu/cm2, respectively, p<0.05). Both, HDLPal and HDLPhl were better substrates for cholesteryl esters transfer protein (CETP) than HDLCtrl (% of transfer, 13.02+/-3.8 and 12.7+/-4.5 vs. 7.8+/-2.7% in 16 h, respectively, p<0.05). HDLPal apo A-I catabolism in vivo, as performed in New Zealand white rabbits by exogenous radiolabeling, was markedly lower than that of HDLPhl and HDLCtrl (fractional catabolic rate, 0.019+/-0.008 vs. 0.030+/-0.005 and 0.047+/-0.003 h-1, respectively, p<0.001), suggesting that negative charge is inversely related to HDL-apo A-I catabolism. CONCLUSIONS: Enrichment with palmitic acid increases the negative electric charge of HDL at physiological pH, contributes to decrease their catabolism, and is associated to an enhanced lipid transfer by CETP that has been related to the atherogenic process.

Animals↗

The use of two in-situ simultaneously-perfused placentas to investigate the effect of carrier proteins on linoleic and palmitic acid transfer in the guinea pig.

By perfusing two placentas concurrently in five animals we have found that linoleic acid is transferred across the guinea-pig placenta more readily than palmitic acid. In the in-situ perfusion this transfer is modulated by the type of albumin used in the perfusate. Removing the associated lipid from the perfusate albumin reduced the amount of non-esterified fatty acid (NEFA) transfer rather than enhanced it. Many researchers use fat free albumin as a protein carrier in in vivo and in-vitro systems, our results indicate that this may not be the most appropriate substance to use.

Animals↗

Glucose-induced preproinsulin gene expression is inhibited by the free fatty acid palmitate.

Prolonged exposure to elevated FFA levels has been shown to induce peripheral insulin resistance and to alter the beta-cell secretory response to glucose. To investigate the effects of FFAs on preproinsulin gene expression, we measured insulin release, cell content, and messenger RNA (mRNA) levels in rat islets after a 24-h exposure to 1 mM palmitate. Insulin release increased at all glucose concentrations studied; in contrast, preproinsulin mRNA levels were specifically reduced by palmitate at high glucose with a decrease in insulin stores, suggesting that palmitate inhibits the glucose-stimulated increase in preproinsulin gene expression. The mechanisms by which palmitate affects preproinsulin gene expression implicate both preproinsulin mRNA stability and transcription, as suggested by an actinomycin D decay assay, quantification of primary preproinsulin transcripts, and transient transfection experiments in Min6 cells. Metabolism of palmitate is not required to obtain these effects, inasmuch as they can be reproduced by 2-bromopalmitate. However, oleate and linoleate did not significantly influence preproinsulin mRNA levels. We conclude that insulin release and preproinsulin gene expression are not coordinately regulated by palmitate and that chronically elevated FFA levels may interfere with beta-cell function and be implicated in the development of noninsulin-dependent diabetes.

Animals↗

Experimental conditions affecting in vitro intestinal incorporation of palmitic acid: a methodological approach.

In the rat, a large number of in vitro studies on intestinal fatty acid uptake have been carried out. However, the results obtained under different experimental conditions are often contradictory. The present work is a critical approach to the experimental aspects which may modify in vitro intestinal uptake of fatty acids. Different kinds of intestinal tissue samples (intact, everted or opened rings) were used. The histological changes and the uptake of palmitic acid were measured for each type of sample under different stirring rates, at different incubation times and with micellar solutions of varying composition. It is concluded that 1) opened rings have the highest absorptive capacity with the lowest dispersion; 2) incubation periods longer than 30 minutes do not result in additional palmitic acid uptake and may lead to severe tissue hypoxia as indicated by extensive vacuolization; 3) stirring rates over 1 cycle/sec do not result in increased PA uptake and cause extensive mucosal disruption, particularly in jejunal samples.

Animals↗

[Diagnosis of chylous ascites with oral administration of 13C-palmitic acid].

12 cases of chylous ascites in PUMC hospital in the recent 30 years were analysed. The etiology of this disease includes non-traumatic (83.3%) and traumatic (16.7%) causes. All the patients received isotope examination or lymphangiography, but only in 8 patients the site of the lesion was found. As these two kinds of examination can only show the lesions in the right and left lumbar lymph trunks, cisterna chyli and throacic duct but not the intestinal lymph truck, the authors set up a method by administering orally 13C-palmitic acid to detect the lesion of intestinal lymph trunk. Palmitic acid is a long-chain fatty acid, which enters directly into the intestinal lymph trunk after absorption. Palmitic acid labelled with 13C could be detected in the ascitic fluid if there is a leak from intestinal lymph trunk. This new method was used to examine a patients who had negative results with isotope examination and lymphangiography; 13C could be detected in the ascitic fluid 30 minutes after oral administration but not in exhaled air. It can be concluded that cyhle leaked into the peritoneal cavity from the intestinal lymph trunk. This method is also of help to determine the possible site of leakage and the degree of obstruction, so it is quite useful for the diagnosis of chylous ascities.

Administration, Oral↗

Replacing dietary palmitic acid with elaidic acid (t-C18:1 delta9) depresses HDL and increases CETP activity in cebus monkeys.

The question whether dietary trans fatty acids affect lipoprotein metabolism similarly to specific saturated fatty acids was investigated in 11 normolipemic cebus monkeys by exchanging 5% dietary energy (%en) between elaidic (t-C18:1 delta9) and palmitic acid (16:0) in two test diets (30%en fat + 100 mg cholesterol/1000 kcal diet) conforming to the American Heart Association (AHA) Step 1 guidelines. These were compared with a normal control diet rich in saturated fat and cholesterol (38%en fat + 180 mg cholesterol/1000 kcal diet). The control diet was fed initially for 14 wk, followed by each of the the two test diets in a crossover design. Plasma lipid concentrations were determined four times between the 6th and 14th wk. Turnover studies (using 125I-HDL and 131I-LDL) were conducted after 9 wk in each dietary period. Relative to the control diet, both test diets significantly reduced plasma total cholesterol (TC), HDL cholesterol (HDL-C) and VLDL plus LDL cholesterol (LDL-C) concentrations; triglyceride (TG) concentrations tended to be lower. However, the trans diet resulted in a significantly greater reduction in HDL-C than the palmitate diet (124 +/- 17, 117 +/- 18 and 106 +/- 13 mg/dL for the control, palmitate and trans diets, respectively). The palmitate diet significantly decreased the TC/HDL-C ratio by 11% when compared with the control diet (1.68 +/- 0.17 vs. 1.89 +/- 0.30), whereas the trans diet had no effect (1.81 +/- 0.20 vs. 1.89 +/- 0.30). Kinetic studies revealed that, relative to the control diet, both test diets significantly lowered the LDL apolipoprotein B (apoB) pool size, principally reflecting an increase in the LDL apoB fractional catabolic rate (FCR) related to the reduced cholesterol intake. Between the two test diets, no significant differences in LDL kinetic parameters were observed. Both test diets significantly decreased HDL apoA1 concentrations in comparison with the control diet, which was partly explained by an increase in the fractional catabolic rate of HDL. Of the two test diets, the trans diet was associated with a 9.5% greater HDL FCR than the palmitate diet (P < 0.08) and a significant increase in plasma cholesteryl ester transfer protein (CETP) activity (% transfer 114 +/- 7 vs. 91 +/- 7; P < 0.03). Thus, palmitic acid- and elaidic acid-rich diets produced identical effects on LDL metabolism in normocholesterolemic cebus monkeys fed diets with low levels of cholesterol, whereas elaidic acid depressed HDL-C, attributable to both increased CETP activity and HDL clearance.

Animals↗

Effect of potassium on utilization of (1-14C)palmitic acid in renal cortex of the rat.

The renal cortex has an unusually high rate of palmitic acid utilization. The uptake, oxidation, and distribution of this fatty acid in the cortex, unlike other tissues, responds to variations in medium and tissue potassium content. This potassium dependency is mimicked and amplified by rubidium. These observations indicate that in the renal cortex, both potassium and rubidium promote the accumulation, oxidation, and esterification of palmitic acid.

Animals↗

Incorporation of palmitic acid in the organ of Corti as revealed by autoradiography.

The sites of incorporation of [3H]palmitic acid perfused through the scala tympani of the guinea pig cochlea were localized autoradiographically. The most active incorporation occurred in the lipid globules of Hensen's cells, followed by the hair cells, myelin of the cochlear nerve and other cells. It is speculated that the lipid globules of Hensen's cells act as a reservoir of the vitamin A esterified by fatty acids.

Animals↗

Effects of gender and pregnancy on hepatocellular uptake of palmitic acid: facilitation by albumin.

The human serum albumin (HSA)-dependent unbound clearance (Clu) of [3H]palmitic acid (PA) by hepatocyte suspensions isolated from immature and mature male and female and pregnant female rats was studied. The Clu values obtained experimentally were compared with the predictions of a noncompartmental diffusion-reaction (Bass-Pond) theory for the cellular uptake of protein-bound ligands. In all groups, as the concentration of HSA (Ca) was increased, there was a striking increase in Clu. These enhancement factors were predicted by the theory. Adult females had higher Clu values at high Ca values than males or immature females. Furthermore, at high Ca values, Clu in pregnant animals was twice as high as in the nonpregnant animals and four times as high as in the aged-matched males. The absolute values of Clu obtained experimentally in both pregnant and nonpregnant females exceeded the maximal predictions of the theory, using reasonable values of all of the parameters. Thus, according to current data on the physicochemical characteristics of the uptake system, the study demonstrates that some specialized process exists to facilitate hepatocellular uptake of fatty acid from albumin, and that it is potentiated by the female sex hormones.

Aging↗

Competency of human-derived Mycobacterium leprae to use palmitic acid in the synthesis of phenolic glycolipid-I and phthiocerol dimycocerosate and to release CO2 in axenic culture.

Insufficient numbers of viable Mycobacterium leprae have hampered metabolic studies using human-derived M. leprae. In this study, sufficient numbers of M. leprae were obtained from an untreated lepromatous patient to titrate the effects of pH on the metabolism of 14C-palmitic acid by M. leprae. Catabolic metabolism (oxidation of 14C-palmitic acid and release of 14CO2) was maximal when M. leprae were incubated at 33 degrees C and suspended in Middlebrook 7H9, ADC supplemented medium that had been buffered to maintain a pH of 4.8. Anabolic metabolism (synthesis of 14C-phenolic glycolipid-I and its precursor, 14C-phthiocerol dimycocerosate) was maximal when the pH was maintained at 6.8.

Adolescent↗

[New procedure in the diagnosis of maldigestion. Studies with 3-H palmitic acid and 14-C tripalmitate in normal subjects and patients with chronic pancreatitis].

3H-palmitic acid and 14C-tripalmitate dissolved in 1.0 g indian corn oil/kg body weight were administered to 29 patients with chronic pancreatitis and 25 control subjects. For the following 8 h 3H and 14C radioactivity in serum lipids and 14CO2 in expired air were measured at 2-hour intervals. Triglyceride absorption was significantly lowered in the pancreatitis group, while the wide dispersion in healthy subjects precluded reliable information on 14C-triglyceride absorption alone in individual cases. When related to palmitic acid absorption, however, reduced triglyceride absorption was much more evident and could be clearly demonstrated in 21 of the 29 patients with chronic pancreatitis. Since none of them had severe pancreatic insufficiency, the new double isotope method presented here appears to offer a very sensitive means of detecting maldigestion or slowed triglyceride hydrolysis. Compared with the 3H-fatty acid absorption, a relatively reduced triglyceride absorption was found in many cases without steatorrhea or otherwise demonstrable pancreatic insufficiency. Additional measurement of 14CO2 appears to be worthless, since in our patients specific activity of 14CO2 in the expired air was not reduced even in cases with mild steatorrhea.

Adult↗

[Metabolism of (1-14C)-palmitic acid in the cat's brain].

Following injection into the cerebral ventricles of conscious cats, (1-14C) palmitic acid was rapidly taken up and incorporated into a variety of brain lipids. The peak of uptake of (1-14C) palmitic acid, about 50% of injected radioactive material, into the brain tissue was obtained within the first 24 h following its administration. Thereafter, the radioactivity slowly decreased reaching the least value by the end of the second week. The most heavily labelled lipids were the phospholipids, while the free fatty acids were appreciably labelled. Small percentage of the radioactive material was found in monoglycerides, diglycerides and triglycerides. The least incorporation was into cholesterol esters.

Animals↗

Incorporation of [1-14C[palmitic acid into neutral lipids and phospholipids of rat cerebral cortex in vitro.

Incorporation of [1-14C]palmitic acid into neutral lipids and phospholipids of rat cerebral cortex was examined in vitro in normal Krebs--Ringer bicarbonate buffer containing 3% (wt/vol) albumin and 0.75 mM palmitic acid. Under standard assay conditions, radioactivity in the triacylglycerol fraction increased rapidly during the first 30 min, and then decreased after 60 min, with corresponding increase in radioactivity in phosphatidyl choline, phosphatidyl ethanolamine, and a fraction of phosphatidyl inositol plus phosphatidyl serine. Diacylglycerol was shown to be an intermediate metabolite. Radioactivity increased in triacylglycerol, and decreased in phosphatidyl choline and phosphatidyl ethanolamine throughout incubation under N2 gas. In the fraction of phosphatidyl inositol plus phosphatidyl serine, radioactivity decreased after 30 min during incubation under N2 gas. A possible acylation--deacylation cycle, in which triacylglycerol could be a source of free fatty acids for phospholipids, is discussed.

Animals↗

Interaction of differently oriented lipids in monolayer: mixed monolayers of 16-(9-anthroyloxy)palmitic acid with phosphatidylcholine and cholesterol.

16-(9-Anthroyloxy)palmitic acid (16-AP) is a bifunctional molecule with carboxyl and 9-anthroyloxy groups attached at both ends of the hydrocarbon chain. At the air-water interface, in a monolayer, the 16-AP molecule has horizontal and vertical orientations, depending on the surface pressure of the monolayer. The miscibilities of 16-AP with dimyristoylphosphatidylcholine (DMPC), cholesterol (CH), and fatty acids in mixed monolayers were evaluated in investigations of monolayer phase transitions. Lipid molecules with flexible hydrocarbon chains, i.e., DMPC and fatty acids, formed homogeneous mixed monolayers with horizontally oriented 16-AP. On the other hand, the rigid molecule, CH, could not accommodate the horizontally oriented 16-AP in a monolayer, and there was a phase separation from 16-AP. In biological and reconstituted membranes, preferential binding of phospholipid to the integral protein and exclusion of cholesterol in close vicinity of the membrane protein have been recognized. On the basis of this work, it can be expected that flexible lipids readily accommodate the rough hydrophobic surface of integral proteins and stabilize the structure of the protein, while rigid lipids such as cholesterol are removed from the immediate environment of the membrane protein, if the protein does not interact specifically with the rigid lipids.

Calorimetry↗

Analysis of organic acids after incubation with (16-2H3)palmitic acid in fibroblasts from patients with mitochondrial beta-oxidation defects.

The analysis of acylcarnitines as products of incubation of intact fibroblasts with isotope-labelled precursors, usually (16-(2)H(3))hexadecanoic acid, is an advanced in vitro method for the study of mitochondrial beta-oxidation defects. We propose a technique for the measurement of the organic acid intermediates after hydrolysis of the acylcarnitines using electron-impact gas chromatography-mass spectrometry. For some mitochondrial beta-oxidation deficiencies, the characteristic profile enables us to approach the diagnosis with clear differentiation.

Acyl-CoA Dehydrogenase↗

Ultrasound accelerated esterification of palmitic acid with vitamin C.

The esterification of palmitic acid with vitamin C in the presence of concentrated sulfuric acid as the solvent and catalyst by means of 25 kHz ultrasonic irradiation to obtain l-ascorbyl 6-palmitate is studied. By using ultrasound the dissolution rate of the reactants can be accelerated greatly, the reaction time of esterification can be reduced from 36 to 2h, and better yield (90-93%) of ester can be given by using 95% concentrated sulfuric acid as the solvent and catalyst, contrast to the yield of 75-85% by using 99% concentrated sulfuric acid without ultrasound. The influence of reaction conditions and ultrasonic parameters to the yield of ascorbyl palmitate are reported.

Ascorbic Acid↗