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[Clinico-experimental long-term study on the combined effect of clofibrin and nicotinic acid on abnormal fatty acid patterns in lipid metabolism disorders in the aged].

Seventy-one patients (forty-two males and twenty-nine females aged forty-five to seventy-six) with disturbances of fat metabolism were treated for three years with clofibric acid and nicotinic acid derivatives. Regular gas chromatographic analyses of the composition of cholesterol ester and triglyceride fatty acids in the serum showed an increase of linoleic, linolenic, arachidonic, and eicosane-pentaenoic acids and a decrease of palmitic, palmitoleic, stearic, oleic, and eicosanetrienic acids during treatment. These changes were far more strongly marked than under monotherapy. Possible causes of the observed changes include selective competitive inhibition of unesterified serum fatty acids, inhibition of lipolysis, effects upon hepatogenic fatty acid metabolism and LCAT, as well as better utilization of alimentary polyunsaturated fatty acids as a result of combination treatment. The increase of polyunsaturated fatty acids and the decrease of saturated and monounsaturated fatty acids with their reciprocal relations to the prostaglandin metabolism may be considered a positive vasoprotective effect which is of particular importance in middle and old age.

Aged↗

Single measurement of serum phospholipid fatty acid as a biomarker of specific fatty acid intake in middle-aged Japanese men.

OBJECTIVE: To assess the utility of serum phospholipid fatty acid (FA) levels as a biochemical indicator of habitual dietary fatty acid intake in Japanese, whose diet is characterized by low fat intake and high intake of n-3 polyunsaturated fatty acids (PUFA) of marine origin. SUBJECTS AND METHODS: Eighty-seven male volunteers from four public health center districts that were part of the Japan Public Health Center based Prospective Study (JPHC Study) cohort I, were included in this study. Habitual intake of fatty acid was obtained by 7 day weighed dietary records four times (in one area only twice) in 1994--1995. Blood was collected twice, in February and August of the same year, and the composition of FA in serum phospholipid was analyzed by gas chromatography. The correlation coefficient between serum phospholipid FA levels and fatty acid intake was calculated. RESULTS: High correlations were observed for eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA), which are marine origin n-3 PUFA (r=0.75, 0.49, 0.50, respectively). No significant correlation was observed for saturated fatty acid (SFA), although the monounsaturated fatty acid (MUFA), palmitoleic acid and oleic acid intake were moderately correlated (r=0.22, 0.35, respectively). The correlations for EPA, DPA and DHA were similar in both samples collected in February and August. CONCLUSIONS: These data suggest that in populations with a high and stable over time intake of n-3 PUFA of marine origin, a single measurement of serum phospholipids reflects the ranking of habitual intake of marine origin n-3 PUFA.

Biomarkers↗

[A new method of evaluating the utilization of nutrients (carbohydrates, amino acids and fatty acids) on the plastic and energy goals in the animal body].

With use of a new method, based on detection in blood serum of radioactivity of water, formed from tritium marked precursors--glucose, amino acids (valine, serine, histidine) and palmitine acid--their distribution on oxidizing and anabolic ways of metabolism was determined. The work was carried out on laboratory rats. In young pubertal rats the ratio of flows on these ways for glucose was found equal 2.83, i.e. it in a greater degree was used as energy substratum. On the contrary, for palmitine acid this ratio was equal 0.10--it was comprised in a plastic material of organism in a greater degree. For serine, histidine and valine it is equal 0.34, 0.71 and 0.46, accordingly. In growing rats the distribution of flows was shifted aside of anabolic way: the ratio of flows is equal 0.19; in old rats--aside of oxidizing: a ratio of flows is equal 0.71.

Amino Acids↗

Rapid uptake and esterification of arachidonic acid and other fatty acids by microfilariae of Brugia malayi.

We have examined the differential incorporation and esterification of exogenous fatty acids by microfilariae of the human filarial parasite Brugia malayi. Microfilariae incubated with 2 nM [3H]arachidonic acid over 1 h rapidly took up this fatty acid. Palmitic, oleic and linoleic acids were also incorporated by parasites. In contrast to these other fatty acids, little incorporated arachidonic acid remained as free fatty acid within microfilariae. Arachidonate was rapidly esterified into phospholipids, with 66% of incorporated arachidonate esterified into phospholipids at 1 min. Esterification of other fatty acids into phospholipids was quantitatively lesser and occurred into phosphatidylcholine and phosphatidylethanolamine. Arachidonate was preferentially esterified into phosphatidylinositol, which constituted only 10% of the total parasite phospholipid pool, and into phosphatidylcholine. By 1 min these two phospholipid classes, respectively, comprised 53% and 43% of [3H]arachidonyl-phospholipids. Neither the microfilarial incorporation of arachidonate nor its esterification into parasite phospholipids could be saturated by noncytotoxic concentrations of up to 600 microM. Microfilariae, which in vivo are exposed to arachidonate in blood, can rapidly, avidly and with high capacity incorporate exogenous arachidonate and esterify it preferentially into specific classes of phospholipids, including phosphatidylinositol. Like many mammalian cells, these phylogenetically distinct metazoan parasites possess efficient means for utilizing host-derived arachidonic acid.

Animals↗

Effects of dietary supplementation with n-3 fatty acids compared with n-6 fatty acids on bronchial asthma.

OBJECTIVE: The effects of perilla seed oil (n-3 fatty acids) on bronchial asthma were compared with the effects of corn oil (n-6 fatty acids) in relation to the pulmonary function and the generation of leukotriene B4 (LTB4) and C4 (LTC4) by leucocytes. METHODS AND SUBJECTS: 14 asthmatic subjects were divided randomly into two groups: one group (7 subjects) consumed perilla seed oil-rich supplementation and the other group (7 subjects) consumed corn oil-rich supplementation for 4 weeks. Generation of LTs by leucocytes and respiratory function were compared between the two groups. RESULTS: The generation of LTB4 and LTC4 by leucocytes tended to increase in subjects (N=7) with corn oil-rich supplementation, and decrease in subjects (N=7) with perilla seed oil-rich supplementation. Significant differences between the two groups were observed in the generation of LTB4 at 2 weeks (p<0.05) and LTC4 at 2 weeks (p<0.05) after dietary supplementation. Significant increases in the value of PEF (p<0.05), FVC (p<0.01), FEV(1.0) (p<0.05) and V(25) (p<0.05) were found in subjects who received perilla seed oil supplementation for 4 weeks. And significant differences in the value of FVC (p<0.05) and FEV(1.0) (p<0.05) were observed between the two groups after 4 weeks of dietary supplementation. CONCLUSION: These results suggest that perilla seed oil-rich supplementation is useful for the treatment of asthma in terms of suppression of LTB4 and LTC4 generation by leucocytes, and improvement of pulmonary function.

Adult↗

A gene encoding a chloroplast omega-3 fatty acid desaturase complements alterations in fatty acid desaturation and chloroplast copy number of the fad7 mutant of Arabidopsis thaliana.

Mutations at the fad7 locus of Arabidopsis thaliana (previously called fadD) cause decreased desaturation of dienoic fatty acids in chloroplast lipids in plants grown at elevated temperatures. This suggested that the fad7 locus encodes a chloroplast omega-3 desaturase that catalyzes the desaturation of lipid-linked 18:2 and 16:2 fatty acids. In order to clone the fad7 gene, it was first genetically mapped relative to the flanking restriction fragment length polymorphism markers 4547 and 2488A on chromosome 3, and yeast artificial chromosomes covering the locus were identified. A putative desaturase cDNA clone that was isolated by low stringency heterologous probing with a cDNA for an endoplasmic reticulum-localized omega-3 desaturase (fad3) hybridized to the yeast artificial chromosomes and could not be resolved from the locus by restriction fragment length polymorphism mapping. Expression of the cDNA in transgenic fad7 mutant plants resulted in restoration of wild type fatty acid composition and suppression of a previously observed effect of the fad7 mutation on chloroplast number, indicating genetic complementation. The structural gene contained seven introns within a coding sequence of 1338 base pairs, which encodes a 446-amino acid polypeptide of 51,172 daltons. The amino-terminal region of the fad7 gene product contained a consensus chloroplast transit peptide. Except for the amino-terminal domain, the deduced amino acid sequence of the fad7 gene product had high homology to the fad3 gene product, indicating that fad7 encodes an omega-3 desaturase and that the two genes arose from a common ancestral gene. There was no apparent effect of growth temperature on the steady-state levels of fad7 mRNA in wild type plants.

Amino Acid Sequence↗

An amino acid substitution in the human intestinal fatty acid binding protein is associated with increased fatty acid binding, increased fat oxidation, and insulin resistance.

The intestinal fatty acid binding protein locus (FABP2) was investigated as a possible genetic factor in determining insulin action in the Pima Indian population. A polymorphism at codon 54 of FABP2 was identified that results in an alanine-encoding allele (frequency 0.71) and a threonine-encoding allele (frequency 0.29). Pimas who were homozygous or heterozygous for the threonine-encoding allele were found to have a higher mean fasting plasma insulin concentration, a lower mean insulin-stimulated glucose uptake rate, a higher mean insulin response to oral glucose and a mixed meal, and a higher mean fat oxidation rate compared with Pimas who were homozygous for the alanine-encoding allele. Since the FABP2 threonine-encoding allele was found to be associated with insulin resistance and increased fat oxidation in vivo, we further analyzed the FABP2 gene products for potential functional differences. Titration microcalorimetry studies with purified recombinant protein showed that the threonine-containing protein had a twofold greater affinity for long-chain fatty acids than the alanine-containing protein. We conclude that the threonine-containing protein may increase absorption and/or processing of dietary fatty acids by the intestine and thereby increase fat oxidation, which has been shown to reduce insulin action.

Adult↗

[Conformational changes of rat liver fatty acid binding protein induced by long chain fatty acid].

Rat Liver fatty acid binding protein (FABP) has been purified to homogeneity by the procedures including Sephadex G-100 and DEAE-cellulose column chromatography. FABP was resolved into two peaks of A and B by DEAE-cellulose column chromatography. Each of these fractions exhibited apparent homogeneity upon polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate with a molecular weight of 16,000 daltons and amino acid analysis of these fractions has revealed that they are identical protein. However, upon isoelectric focusing on polyacrylamide gel, the isoelectric (pl) points were differed considerably showing microheterogeneity. In the present investigation, one isoform (pl = 5.0) of FABP was purified by a successive DEAE-cellulose column chromatography and used for the subsequent experiments of fatty acyl-protein interactions. When the final FABP preparation was partly freed of fatty acids by a mild delipidation technique using Lipidex, the pl shifted toward higher regions of 7.0. However, the pl of the delipidated FABP turned to the original 7.0 by recombining fatty acids. On the other hand, the secondary and tertiary structures were also significantly changed by delipidization, which have been demonstrated by circular dichroism (CD) and nuclear magnetic resonance (1H-NMR) spectroscopy. Furthermore the structural properties of the delipidated FABP also could be restored by recombining fatty acid. These findings suggested that the weakly bound fatty acids are responsible for the functional capacity of the FABP by virtue of changing the protein conformation.

Animals↗

Correlation between the activity of glucosylceramidase and its binding to glucosylceramide-containing liposomes. Role of acidic phospholipids and fatty acids.

Optimal enzymatic hydrolysis of glucosylceramide inserted into liposomes has been obtained when both acidic phospholipids and the appropriate fatty acids were added to glucosylceramide-containing liposomes. In fact, the stimulation of glucosylceramidase by acidic phospholipids was synergistically enhanced by fatty acids, whose effect was dependent upon chain length and increased on unsaturation. By following the partition of glucosylceramidase between the aqueous phase and the liposome-associated state with a flotation procedure, it has been found that phosphatidic acid (PA) and oleic acid (OA), as representatives of acidic phospholipids and activating fatty acids, respectively, were both required not only for optimal glucosylceramidase activity, but also for a tight binding of the enzyme to the liposomes. The binding was significantly less effective in the absence of either PA or OA. In the absence of both PA and OA no physical interaction between the enzyme and the liposomes was observed. Under all conditions, the glucosylceramidase activity directly correlated with the enzyme binding to the substrate-containing liposomes. Additionally, we have obtained evidence that the site(s) of the enzyme involved in the binding to the liposomes is distinct from the catalytic site; in fact, the enzyme could still associate with liposomes containing PA and OA but devoid of glucosylceramide, while it was incapable of binding to glucosylceramide-containing liposomes in the absence of PA and OA. In conclusion, the presence in liposomes of acidic phospholipids together with the appropriate fatty acids plays a key role in promoting the binding of glucosylceramidase. Consequently, when glucosylceramide is also included in the liposomes, its hydrolysis is markedly enhanced by these acidic lipids.

Binding Sites↗

Influence of dietary fatty acid composition and vitamin E on fatty acids and alpha-tocopherol in carp (Cyprinus carpio L.).

This paper examines the effect of different fatty acid composition, together with varying dietary vitamin E contents on fatty acid composition and tocopherol and tocotrienol contents of carp, and compares with other species of fish and the storage quality of the fish produced. Dietary fatty acid composition affected fatty acid patterns of tissues of carp significantly, but this was correlated with the fat content of the diet. Varying the vitamin E content results in no or only very slight variations in the fatty acid pattern of the tissues. The content of alpha-tocopherol in the tissues was affected significantly by dietary alpha-tocopheryl acetate content and by dietary fat source. There was a negative correlation between (n-3) fatty acids and the storage of alpha-tocopherol, while this was positively correlated with (n-6) fatty acids. The content of alpha-tocopherol in fish affected the stability during storage with the content of malondialdehyde used as a parameter for measuring the extent of fat oxidation. Other relationships, e.g. with sensory qualities and S-containing substances in the edible part of the fish, are discussed.

Adipose Tissue↗