Choline Supplementation With a Structured Lipid in Children With Cystic Fibrosis: A Randomized Placebo-Controlled Trial.

BACKGROUND: Choline depletion is seen in cystic fibrosis (CF) and pancreatic insufficiency in spite of enzyme treatment and may result in liver, fatty acid, and muscle abnormalities. This study evaluated the efficacy and safety of an easily absorbed choline-rich structured lipid (LYM-X-SORB™ [LXS]) to improve choline status. METHODS: Children with CF and pancreatic insufficiency were randomized to LXS or placebo in a 12-month double blind trial. Dietary choline intake, plasma cholines, plasma and fecal phospholipids, coefficient of fat absorption, pulmonary function, growth status, body composition, and safety measures were assessed. Magnetic resonance spectroscopy for calf muscle choline and liver fat were assessed in a subgroup and compared with a healthy comparison group matched for age, sex, and body size. RESULTS: A total of 110 subjects were enrolled (age 10.4 ±â€Š3.0 years). Baseline dietary choline, 88% recommended, increased 3-fold in the LXS group. Plasma choline, betaine, and dimethylglycine increased in the LXS but not placebo (P = 0.007). Plasma lysophosphatidylcholine and phosphatidylcholine increased, and fecal phosphatidylcholine/phosphatidylethanolamine ratio decreased (P ≤ 0.05) in LXS only, accompanied by a 6% coefficient of fat absorption increase (P = 0.001). Children with CF had higher liver fat than healthy children and depleted calf muscle choline at baseline. Muscle choline concentration increased in LXS and was associated with improvement in plasma choline status. No relevant changes in safety measures were evident. CONCLUSIONS: LXS had improved choline intake, plasma choline status, and muscle choline stores compared with placebo group. The choline-rich supplement was safe, accepted by participants, and improved choline status in children with CF.

Application of imaging mass spectrometry for the analysis of Oryza sativa rice.

Rice is one of the most important food crops in the world and new varieties have been bred for specific purposes, such as the development of drought-resistance, or the enrichment of functional food factors. The localization and composition of metabolites in such new varieties must be investigated because all artificial interventions are expected to change the metabolites of rice. Imaging mass spectrometry using matrix-assisted laser desorption/ionization (MALDI-IMS) is a suitable tool for investigating the localization and composition of metabolites; however, suitable methodologies for the MALDI-IMS analysis of rice have not yet been established. In this study, we optimized the methods for analyzing rice grains by MALDI-IMS using adhesive film and found the characteristic distribution of metabolites in rice. Lysophosphatidylcholine (LPC) was localized in the endosperm. Phosphatidylcholine (PC), gamma-oryzanol and phytic acid were localized in the bran (germ and seed coat), and alpha-tocopherol was distributed in the germ (especially in the scutellum). In addition, MALDI-IMS revealed the LPC and PC composition of the rice samples. The LPC composition, LPC (1-acyl 16:0), LPC (1-acyl 18:2), LPC (1-acyl 18:1) and LPC (1-acyl 18:0), was 59.4 +/- 4.5%, 19.6 +/- 2.5%, 14.2 +/- 4.5% and 6.8 +/- 1.4%. The PC composition, PC (diacyl 16:0/18:2), PC (diacyl 16:0/18:1), PC (diacyl 18:1/18:3), PC (diacyl 18:1/18:2) and PC (diacyl 18:1/18:2), was 19.6 +/- 1.0%, 21.0 +/- 1.0%, 15.0 +/- 1.4%, 26.7 +/- 0.7% and 17.8 +/- 1.9%. This approach can be applied to the assessment of metabolites not only in rice, but also in other foods for which the preparation of sections is a challenging task.

[Antioxidants inhibit the oxidative modification of high density lipoproteins].

OBJECTIVE: To study the role and mechanism of antioxidants on inhibiting oxidative modification of high density lipoproteins (HDL). METHODS: Freshly prepared human plasma HDL was treated by incubation with copper ion, hyperchlorite or arterial wall cells. Compared to control, the test groups were treated with addition of different concentration of butylhydroxytoluene (BHT), vitamin C and vitamin E. Then, the relative electrophoretic mobility (REM), thiobarbituric acid-reactive substances (TBARS), ratio of lysolecithin to lecithin (LPC/PC), and lipoprotein moieties were investigated. RESULTS: BHT, vitamin C and vitamin E can significantly inhibit the increasing REM, TBARS, LPC/PC ratio and lipoprotein variation that induced by copper ion and hyperchlorite and arterial wall cells. But these antioxidants act on different manner. CONCLUSION: BHT, vitamin C and vitamin E can inhibit the oxidative modification of HDL and hence could be potential nutrients to prevent atherosclerosis.

Bitter gourd suppresses lipopolysaccharide-induced inflammatory responses.

Bitter gourd ( Momordica charantia L.) is a popular tropical vegetable in Asian countries. Previously it was shown that bitter gourd placenta extract suppressed lipopolysaccharide (LPS)-induced TNFalpha production in RAW 264.7 macrophage-like cells. Here it is shown that the butanol-soluble fraction of bitter gourd placenta extract strongly suppresses LPS-induced TNFalpha production in RAW 264.7 cells. Gene expression analysis using a fibrous DNA microarray showed that the bitter gourd butanol fraction suppressed expression of various LPS-induced inflammatory genes, such as those for TNF, IL1alpha, IL1beta, G1p2, and Ccl5. The butanol fraction significantly suppressed NFkappaB DNA binding activity and phosphorylation of p38, JNK, and ERK MAPKs. Components in the active fraction from bitter gourd were identified as 1-alpha-linolenoyl-lysophosphatidylcholine (LPC), 2-alpha-linolenoyl-LPC, 1-lynoleoyl-LPC, and 2-linoleoyl-LPC. Purified 1-alpha-linolenoyl-LPC and 1-linoleoyl-LPC suppressed the LPS-induced TNFalpha production of RAW 264.7 cells at a concentration of 10 microg/mL.

Insecticidal components from field pea extracts: soyasaponins and lysolecithins.

Extracts from field peas (Pisum sativum L.) have previously been shown to have a utility to control insect pests. To identify potentially new bioinsecticides in field crops, we describe the fractionation of impure extracts (C8 extracts) derived from protein-rich fractions of commercial pea flour. The activity of separated fractions was determined by a flour disk antifeedant bioassay with the rice weevil [Sitophilus oryzae (L.)], an insect pest of stored products. Bioassay-guided fractionation showed that the triterpenoid saponin fractions were partly responsible for the antifeedant activity of C8 extracts. Soyasaponin I (soyasaponin Bb), isolated from peas and soybeans, and mixtures of soyasaponins, comprised of soyasaponins I-III and isolated from soybeans, were inactive antifeedants, but dehydrosoyasaponin I (the C-22 ketone derivative of soyasaponin I), a minor component found in C8 extracts, was shown to be an active component. Dehydrosoyasaponin I (soyasaponin Be) and soyasaponin VI (soyasaponin betag) coeluted under conditions of silica gel thin-layer chromatography and C18 high-performance liquid chromatography. However, dehydrosoyasaponin I could be isolated from saponin-enriched fractions with a reversed phase column of styrene/divinylbenzene operated at alkaline pH. Phospholipids of the lysolecithin type were also identified in saponin fractions of C8 extracts from peas. Three of the lysolecithins were inactive alone against rice weevils, but mixtures of these phospholipids enhanced the insecticidal activity of dehydrosoyasaponin I.

Lipid analysis of bronchoalveolar lavage fluid (BAL) by MALDI-TOF mass spectrometry and 31P NMR spectroscopy.

Despite the high clinical relevance, only the cellular moiety of bronchoalveolar lavage (BAL) has been intensively investigated and is used for diagnosis purposes. On the other hand, the cell-free fluid is, by far, less characterized. Although this fluid represents a relatively simple mixture of only a few different phospholipids (mainly phosphatidylcholine, phosphatidylglycerol and cholesterol), methods for the routine analysis of these fluids are still lacking. In the present investigation we have applied, for the first time, MALDI-TOF mass spectrometry, as well as 31P NMR spectroscopy to the analysis of organic extracts of bronchoalveolar lavage fluids. BAL from different mammals (rat, minipig, rabbit and man) were investigated and, for means of comparison, organic extracts of lung tissue were also examined. Both applied methods provide fast and reliable information on the lipid composition of the bronchoalveolar lavage. However, despite of its comparably low sensitivity, 31P NMR spectroscopy detects all phospholipid species in a single experiment and with the same sensitivity, whereas MALDI-TOF fails in the detection of phosphatidylethanolamine in the presence of higher quantities of phosphatidylcholine. In contrast, MALDI-TOF mass spectrometry is more suitable for the detection of cholesterol and the determination of the fatty acid composition of the individual phospholipids, especially lysolipids. It will be shown that all BALs exhibit significant, species-dependent differences that mainly concern the content of phosphatidylglycerol and lyso-phosphatidylcholine. It is concluded that both methods are suitable tools in lipid research due to the (in comparison to alternative methods) simplicity of performance.

Dietary phospholipid-dependent reductions in gene expression and activity of liver enzymes in fatty acid synthesis in fasted-refed rats.

The effects of dietary soybean phospholipid, its hydrogenation product and safflower phospholipid on gene expression and the activity of hepatic enzymes in fatty acid biosynthesis were examined in fasted-refed rats. Phospholipid composition of soybean phospholipid and its hydrogenation product were the same, but the hydrogenation product contained negligible amounts of unsaturated fatty acids. Among phospholipid classes, lysophosphatidylcholine and phosphatidylinositol proportions were slightly higher in safflower phospholipid than in soybean phospholipid or its hydrogenation product. Rats were fasted for 2 d and refed a fat-free diet or a diet containing 4% fatty acids either as soybean oil or various phospholipid preparations for 3 d. Compared to the fat-free diet, the soybean oil diet only slightly decreased specific, but not total hepatic fatty acid synthetase and malic enzyme activity, and it was totally ineffective in modulating glucose 6-phosphate dehydrogenase and pyruvate kinase activity under our experimental conditions. The diets containing phospholipids, however, markedly decreased the activity of these enzymes. The extent of reduction was somewhat attenuated with hydrogenated soybean phospholipid as compared with soybean and safflower phospholipids. Dot and Northern blot hybridization using specific cDNA probes showed that, compared to a fat-free diet, diets containing phospholipids profoundly decreased the hepatic mRNA levels of enzymes in fatty acid synthesis. Soybean oil, however, only marginally affected these parameters. Hepatic mRNA levels for enzymes correlated well with enzyme activity. Dietary phospholipids therefore appear to have decreased enzyme activity in fatty acid synthesis primarily by suppressing the mRNA levels of these enzymes. Compared to soybean oil, hydrogenated soybean phospholipid is still effective in decreasing the activity and mRNA level of enzymes in fatty acid synthesis. Therefore, it is difficult to ascribe the potent physiological activity of phospholipid in reducing fatty acid synthesis entirely to polyunsaturated fatty acid moiety.

Phospholipase A2 stimulates rat gastric epithelial cell line (RGM-1) migration.

OBJECTIVE AND METHODS: The effect of phospholipase A2 (PLA2) and its phospholipid metabolites on gastric epithelial migration was examined using an in vitro wounding model of confluent monolayers of rat gastric epithelial cell line RGM-1. RESULTS: Lysophosphatidylcholine (lysoPC) (0.01 100 ng/ml) as well as PLA2 (0.01-100 mU/ml) dose-dependently increased the cell migration. Lysophosphatidic acid (10 ng/ml) also increased the migration, but no significant increase in migration was observed when stimulated by lysophosphatidylethanolamine (10 ng/ml) or lysophosphatidylserine (10 ng/ml). Addition of 4-bromophenacyl bromide (BPB), a PLA2 inhibitor, completely blocked the effect of PLA2. However, addition of piroxicam (a cyclooxygenase inhibitor) or nordihydroguaiaretic acid (a lipoxygenase inhibitor) had no significant effect. Combination of PLA2 (10 mU/ml) with lysoPC (10 ng/ml) had no additive effect on migration. Moreover, lysoPC levels were increased in the cells after incubation with PLA2 (10 mU/ml). After pretreatment of RGM-1 cells with replication-inhibiting doses of mitomycin C. PLA2 and lysoPC still increased the cell migration. CONCLUSIONS: These data suggest that PLA2 may, independently of proliferation, increase gastric epithelial migration mainly via lysoPC production.

Restorative effect of shosaikoto (kampo medicine) on diminution of nitric oxide synthesis in murine peritoneal macrophages induced by hypercholesterolemia.

Macrophages play important roles both in immune response and in lipid metabolism and contribute to the development of atherosclerosis. To clarify the mechanism by which Shosaikoto, a Kampo medicine, shows anti-atherosclerotic action, we studied its effect on macrophage function. The production of nitric oxide (NO), prostaglandin E2 and interleukin 1 by macrophages in mice was reduced by feeding of a cholesterol-enriched diet, and the reduced production was observed 1 week after the beginning of cholesterol feeding. Furthermore, although oxidized low density lipoprotein (LDL) and lysophosphatidylcholine (LPC) reduced NO production, macrophages prepared from mice treated with Shosaikoto at a dose of 1.2 g/kg/d restored the reduced NO production by them as well as by hypercholesterolemia. When the content of LPC was measured, no difference was observed between mice fed a cholesterol-enriched diet in the presence or absence of Shosaikoto treatment, suggesting that the restorative effect of Shosaikoto is not due to the inhibition of LPC production or accumulation. Conclusively, Shosaikoto prevents the modification of macrophage function induced by atherogenic factors, which is probably linked to its displayed anti-atherosclerotic action.

[Lipid composition and fertility of human ejaculate]. / Lipidovyi sklad ta fertyl'na spromozhnist' liuds'koho eiakuliatu.

The morpho-functional indexes of ejaculates from 35 healthy and 145 infertile men were studied. The depression of functional sperm activity was found in the majority of infertile men. Phospholipid analysis of whole ejaculates from 12 healthy and 35 infertile subjects was performed. It was shown that inorganic phosphorus of total phospholipids decreased in ejaculates of men with secretory infertility. Lyso-phosphatidyl choline (LPC) was not detected in ejaculates of men with relative infertility. The amount of lyso-phosphatidyl ethanolamine (lyso-PE) and sphingomyelin in whole ejaculates of those patients decreased. The levels of LPC and lyso-PE decreased in ejaculates of persons with associated infertility. The level of phosphatidyl inositol, LPC and PE decreased in ejaculates of men with secretory infertility. The gas-lipid chromatography analysis of fatty acids composition of whole ejaculates of the infertile men showed significant changes in quantity of fatty acids. The quantity of docosahexaenoic acid in ejaculates of infertile men decreased and positively correlated with motility of spermatozoa.

The redistribution of bulk aqueous phase phospholipids during thermal stressing of phospholipid-stabilized emulsions.

The mechanism of the stabilization of triglyceride emulsions by phospholipids has been studied using an HPLC-FID method to determine phosphatidylcholine, lyso-phosphatidylcholine and phosphatidylethanolamine in the oil and aqueous phases of a model emulsion consisting of soybean oil 20 g, egg phospholipid 1.2 g, glycerol 2.25 g and water to 100 mL. It was shown that, on heat sterilization of the emulsions, the phospholipids rapidly relocate from the aqueous phase to the oil phase. It is suggested that the phospholipids concentrate in the oil/water meso phase, forming a cubic liquid crystalline phase, the bulk of which is converted to a lamellar phase on cooling, and that this organization of interfacial material accounts for the enhanced stability of phospholipid emulsions after heat sterilization.

Duodenogastric reflux in patients with upper abdominal complaints or gastric ulcer with particular reference to reflux-associated gastritis.

The aim of this survey was to examine the incidence of duodenogastric reflux in patients with abdominal complaints and the relations between the nature and extent of reflux abdominal complaints, the use of drugs, smoking, the drinking of coffee and alcohol and histological changes in the gastric mucosa. A comparison was also made between gastric ulcer patients and patients with upper abdominal complaints with respect to the nature and extent of reflux. The patients examined included 107 with abdominal complaints and 33 with a gastric ulcer. Gastroscopy was performed, followed by determination of intragastric bile acids and lysolecithin and a duodenogastric isotope reflux examination using technetium-99m-diethyliminodiacetic acid (Tc-99m HIDA). Intragastric bile acid concentrations in the patients with upper abdominal complaints were in the range 7-21,458 mumol/l (mean 964 +/- 2342 mumol/l) and lysolecithin concentrations in the range 0-1992 mumol/l (mean 70 +/- 273 mumol/l). Isotope reflux was observed in 48% of the patients, the reflux index varying in the range 0-70% (mean 4 +/- 9%). The patients suffered more frequently from nausea, epigastric fullness and flatulence with increasing reflux, as assessed by the various methods used here, but only the increase in epigastric fullness symptoms with rising intragastric bile acid concentrations was statistically significant (p less than 0.05). Similarly the various measures of reflux were higher in those patients taking anticholinergic, psychotherapeutic or cardiovascular drugs, antacids or metoclopramide than in the patients not taking the respective drugs, although the only statistically significant increases were in intragastric bile acids among the users of antacids and metoclopramide (p less than 0.01 and p less than 0.05, respectively) and the increase in lysolecithin concentrations among those taking metoclopramide (p less than 0.05). Those abstaining from alcohol had an intragastric bile acid concentration over 1000 mumol/l significantly more often than those who drank alcohol (p less than 0.05), but smoking and the drinking of coffee showed no significant correlation with duodenogastric reflux. The body gastritis score increased significantly with the extent of isotope reflux and the concentrations of intragastric bile acids (p less than 0.05 and p less than 0.01, respectively), and the latter also showed a significant correlation with serum gastrin (p less than 0.05). No significant relationship could be detected between intragastric lysolecithin concentrations and the gastritis score.(ABSTRACT TRUNCATED AT 400 WORDS)

Lipd profiles of plasma lipoproteins of fasted and fed normal and choline-deficient rats.

Three major density classes of lipoproteins and a residual protein (d greater than 1.21) were isolated by ultracentrifugation from plasma of fasted, fed normal, and choline-deficient rats. Lipid extracts were obtained from total plasma and the various density classes of lipoproteins, and each extract was examined in detail by thin layer and gas chromatographies. The results indicated essentially identical compositions of molecular species of phosphatidyl choline, which suggested their rapid equilibration among the different plasma lipoprotein classes. In contrast, the molecular species of the triacylglycerols and cholesteryl esters showed significant differences among the chylomicrons, very low and low, and high density lipoproteins, which excluded the possibility of their ready equilibration in vivo. Omission of choline from diet resulted in a sharp and statistically significant decrease in all lipid components of the very low and low density lipoproteins with 2 days. After 10 days of choline deficiency, the lipid levels of chylomicrons and very low and low density lipoproteins were ca. one-half the levels found in the choline supplemented animals, and there were discernible distortions in their lipid composition. Reintroduction of choline led to a prompt return to normal levels and lipid composition of both chylomicron and very low and low density lipoprotein fractions. The lack of equilibration of the triacylglycerols among the lipoprotein classes under normal conditions and in choline deficiency demonstrates an as yet unrecgnized source of compartmentation of plasma lipids.