Showing papers in "Lipids in 2009"
TL;DR: The findings provide support for unifying the disparate markers of MetS and for the proposed intimate connection with dietary carbohydrate in a 12-week study comparing two hypocaloric diets.
Abstract: We recently proposed that the biological markers improved by carbohydrate restriction were precisely those that define the metabolic syndrome (MetS), and that the common thread was regulation of insulin as a control element. We specifically tested the idea with a 12-week study comparing two hypocaloric diets (~1,500 kcal): a carbohydrate-restricted diet (CRD) (%carbohydrate:fat:protein = 12:59:28) and a low-fat diet (LFD) (56:24:20) in 40 subjects with atherogenic dyslipidemia. Both interventions led to improvements in several metabolic markers, but subjects following the CRD had consistently reduced glucose (−12%) and insulin (−50%) concentrations, insulin sensitivity (−55%), weight loss (−10%), decreased adiposity (−14%), and more favorable triacylglycerol (TAG) (−51%), HDL-C (13%) and total cholesterol/HDL-C ratio (−14%) responses. In addition to these markers for MetS, the CRD subjects showed more favorable responses to alternative indicators of cardiovascular risk: postprandial lipemia (−47%), the Apo B/Apo A-1 ratio (−16%), and LDL particle distribution. Despite a threefold higher intake of dietary saturated fat during the CRD, saturated fatty acids in TAG and cholesteryl ester were significantly decreased, as was palmitoleic acid (16:1n-7), an endogenous marker of lipogenesis, compared to subjects consuming the LFD. Serum retinol binding protein 4 has been linked to insulin-resistant states, and only the CRD decreased this marker (−20%). The findings provide support for unifying the disparate markers of MetS and for the proposed intimate connection with dietary carbohydrate. The results support the use of dietary carbohydrate restriction as an effective approach to improve features of MetS and cardiovascular risk.
362 citations
TL;DR: The multiple actions of U18666A have enabled major discoveries in lipid research and contributed to understanding the pathophysiology of multiple diseases as well as providing animal models for two important disorders: petite mal (absence) epilepsy and cataracts.
Abstract: The multiple actions of U18666A have enabled major discoveries in lipid research and contributed to understanding the pathophysiology of multiple diseases. This review describes these advances and the utility of U18666A as a tool in lipid research. Harry Rudney's recognition that U18666A inhibited oxidosqualene cyclase led him to discover a pathway for formation of polar sterols that he proved to be important regulators of 3-hydroxy-3-methyl-glutaryl coenzyme A reductase. Laura Liscum's recognition that U18666A inhibited the egress of cholesterol from late endosomes and lysosomes led to greatly improved perspective on the major pathways of intracellular cholesterol trafficking. The inhibition of cholesterol trafficking by U18666A mimicked the loss of functional Niemann-Pick type C protein responsible for NPC disease and thus provided a model for this disorder. U18666A subsequently became a tool for assessing the importance of molecular trafficking through the lysosomal pathway in other conditions such as atherosclerosis, Alzheimer's disease, and prion infections. U18666A also provided animal models for two important disorders: petite mal (absence) epilepsy and cataracts. This was the first chronic model of absence epilepsy. U18666A is also being used to address the role of oxidative stress in apoptosis. How can one molecule have so many effects? Perhaps because of its structure as an amphipathic cationic amine it can interact and inhibit diverse proteins. Restricting the availability of cholesterol for membrane formation through inhibition of cholesterol synthesis and intracellular trafficking could also be a mechanism for broadly affecting many processes. Another possibility is that through intercalation into membrane U18666A can alter membrane order and therefore the function of resident proteins. The similarity of the effects of natural and enantiomeric U18666A on cells and the capacity of intercalated U18666A to increase membrane order are arguments in favor of this possibility.
205 citations
TL;DR: The determination of the enzymatic activity of the six lipoxygenases found in the genome of the model plant Arabidopsis thaliana correlated with that predicted by their phylogenetic relationship to other biochemically-characterized plant lip oxygengenases.
Abstract: Lipoxygenases (LOX) catalyze the oxygenation of polyunsaturated fatty acids, the first step in the biosynthesis of a large group of biologically active fatty acid metabolites collectively named oxylipins. In the present study we report the characterization of the enzymatic activity of the six lipoxygenases found in the genome of the model plant Arabidopsis thaliana. Recombinant expressed AtLOX-1 and AtLOX-5 had comparable oxygenase activity with either linoleic acid or linolenic acid. AtLOX-2, AtLOX-3, AtLOX-4 and AtLOX-6 displayed a selective oxygenation of linolenic acid. Analyses by high-performance liquid chromatography and gas chromatography-mass spectrometry demonstrated that AtLOX-1 and AtLOX-5 are 9S-lipoxygenases, and AtLOX-2, AtLOX-3, AtLOX-4 and AtLOX-6 are 13S-lipoxygenases. None of the enzymes had dual positional specificity. The determined activities correlated with that predicted by their phylogenetic relationship to other biochemically-characterized plant lipoxygenases.
196 citations
TL;DR: It appears that dietetic supplementation with coconut oil does not cause dyslipidemia and seems to promote a reduction in abdominal obesity.
Abstract: The effects of dietary supplementation with coconut oil on the biochemical and anthropometric profiles of women presenting waist circumferences (WC) >88 cm (abdominal obesity) were investigated. The randomised, double-blind, clinical trial involved 40 women aged 20–40 years. Groups received daily dietary supplements comprising 30 mL of either soy bean oil (group S; n = 20) or coconut oil (group C; n = 20) over a 12-week period, during which all subjects were instructed to follow a balanced hypocaloric diet and to walk for 50 min per day. Data were collected 1 week before (T1) and 1 week after (T2) dietary intervention. Energy intake and amount of carbohydrate ingested by both groups diminished over the trial, whereas the consumption of protein and fibre increased and lipid ingestion remained unchanged. At T1 there were no differences in biochemical or anthropometric characteristics between the groups, whereas at T2 group C presented a higher level of HDL (48.7 ± 2.4 vs. 45.00 ± 5.6; P = 0.01) and a lower LDL:HDL ratio (2.41 ± 0.8 vs. 3.1 ± 0.8; P = 0.04). Reductions in BMI were observed in both groups at T2 (P < 0.05), but only group C exhibited a reduction in WC (P = 0.005). Group S presented an increase (P < 0.05) in total cholesterol, LDL and LDL:HDL ratio, whilst HDL diminished (P = 0.03). Such alterations were not observed in group C. It appears that dietetic supplementation with coconut oil does not cause dyslipidemia and seems to promote a reduction in abdominal obesity.
153 citations
TL;DR: Targeted mutagenesis of one gene of this synthase was conducted to confirm PUFA synthase function and determine its metabolic necessity, and the resulting mutants were auxotrophic and required supplementation with PUFAs.
Abstract: Schizochytrium produces long chain polyunsaturated fatty acids (PUFAs) via a PUFA synthase. Targeted mutagenesis of one gene of this synthase was conducted to confirm PUFA synthase function and determine its metabolic necessity. The resulting mutants were auxotrophic and required supplementation with PUFAs. In vivo labeling experiments with radioactive fatty acids demonstrated the presence of several elongase and desaturase activities associated with the standard pathway of PUFA synthesis. However, this system was missing a critical Δ12 desaturase activity and was therefore not capable of synthesizing PUFAs from the 16- or 18-carbon saturated fatty acid products of the fatty acid synthase. Because Schizochytrium uses a PUFA synthase system for the production of PUFAs, the existence of a partial desaturase-elongase system (if not a simple vestige) is suggested to be either a scavenging mechanism for intermediate fatty acids prematurely released by the PUFA synthase or for PUFAs found in the organism’s native environment.
143 citations
TL;DR: The data suggested that tocotrienols are better anti-inflammatory agents than α-tocopherol and the most effective form is δ-tocotrienol.
Abstract: Tocotrienols are powerful chain breaking antioxidant Moreover, they are now known to exhibit various non-antioxidant properties such as anti-cancer, neuroprotective and hypocholesterolemic functions This study was undertaken to investigate the anti-inflammatory effects of tocotrienol-rich fraction (TRF) and individual tocotrienol isoforms namely δ-, γ-, and α-tocotrienol on lipopolysaccharide-stimulated RAW2647 macrophages The widely studied vitamin E form, α-tocopherol, was used as comparison Stimulation of RAW2647 with lipopolysaccharide induced the release of various inflammatory markers 10 μg/ml of TRF and all tocotrienol isoforms significantly inhibited the production of interleukin-6 and nitric oxide However, only α-tocotrienol demonstrated a significant effect in lowering tumor necrosis factor-α production Besides, TRF and all tocotrienol isoforms except γ-tocotrienol reduced prostaglandin E2 release It was accompanied by the down-regulation of cyclooxygenase-2 gene expression by all vitamin E forms except α-tocopherol Collectively, the data suggested that tocotrienols are better anti-inflammatory agents than α-tocopherol and the most effective form is δ-tocotrienol
109 citations
TL;DR: Wujin pigs possessed higher mRNA abundance, protein expression or enzyme activities of anabolism, fatty acid transportation and desaturation, and lower catabolism compared with Landrace pigs, suggesting the mechanism of higher IMF content in fatty pigs may be due to the higher capacity of lipogenesis and fatty acid Transportation and the lower capacity of Lipolysis.
Abstract: Intramuscular fat (IMF) content affects meat quality and varies in different pig breeds. However, the underlying mechanisms of different IMF depositions in different genetic backgrounds of pigs have not been fully elucidated as yet. Lipid metabolism theoretically contributes to the variation of IMF content. The expression levels of genes and proteins as well as enzyme activities implicated in muscle lipid metabolism were investigated, which included lipogenetic genes (SREBP-1c and FAS), fatty acid transporting genes (H-FABP and A-FABP), fatty acid oxidative gene (CPT-1B) and lipolytic genes (ATGL and HSL) as well as the desaturated fatty acid gene (SCD). Longissimus muscle samples were collected from fatty Wujin pigs and lean Landrace pigs. Results showed that the average daily gain of Wujin pigs was lower than that of Landrace pigs. Wujin pigs had greater adipocyte diameter, IMF content and PUFA percentage than that of Landrace pigs. Compared with Landrace pigs, Wujin pigs exhibited higher expression levels, both in mRNA and protein, of FAS, SREBP-1c, SCD, A-FABP and H-FABP genes and lower expression levels of CPT-1B, HSL and ATGL genes. Overall, Wujin pigs possessed higher mRNA abundance, protein expression or enzyme activities of anabolism, fatty acid transportation and desaturation, and lower catabolism. Therefore, the mechanism of higher IMF content in fatty pigs may be due to the higher capacity of lipogenesis and fatty acid transportation and the lower capacity of lipolysis.
101 citations
TL;DR: These studies show 9-NO2-OA and 10-NO 2-OA as endogenous nitrated fatty acids in human plasma in the pM range; HPLC is recommendable as a sample clean-up step for reliable quantification of nitro-oleic acids by GC–MS/MS.
Abstract: First studies on the occurrence of nitrated fatty acids in plasma of healthy subjects revealed basal concentrations of 600 nM for free/nonesterified nitro-oleic acid (NO(2)-OA) as measured by liquid chromatography tandem mass spectrometry (LC-MS/MS). We recently showed by a gas chromatography tandem mass spectrometry (GC-MS/MS) method the physiological occurrence of two isomers, i.e., 9-NO(2)-OA and 10-NO(2)-OA, at mean basal plasma concentrations of 880 and 940 pM, respectively. In consideration of this large discrepancy we modified our originally reported method by replacing solid-phase extraction (SPE) by solvent extraction with ethyl acetate and by omitting the high-performance liquid chromatography (HPLC) step for a more direct detection and with the potential for lipidomics studies. Intra-assay imprecision and accuracy of the modified method in human plasma were 1-34% and 91-221%, respectively, for added NO(2)-OA concentrations in the range 0-3,000 pM. This method provided basal plasma concentrations of 306 +/- 44 pM for 9-NO(2)-OA and 316 +/- 33 pM for 10-NO(2)-OA in 15 healthy subjects. Nitro-arachidonic acid and nitro-linolenic acid were not detectable in the plasma samples. In summary, our studies show 9-NO(2)-OA and 10-NO(2)-OA as endogenous nitrated fatty acids in human plasma in the pM range; HPLC is recommendable as a sample clean-up step for reliable quantification of nitro-oleic acids by GC-MS/MS.
82 citations
TL;DR: AKE showed excellent antibacterial activities against drug-susceptible and -resistant Propionibacterium acnes and Staphylococcus epidermidis, which are acne-causing bacteria, and is suggested that AKE may be an attractive candidate for promoting skin health.
Abstract: Since acne vulgaris is the combined result of a bacterial infection and the inflammatory response to that infection, we examined whether Abies koreana essential oil (AKE) possessed anti-inflammatory and antibacterial activities against skin pathogens. In this study, AKE showed excellent antibacterial activities against drug-susceptible and -resistant Propionibacterium acnes and Staphylococcus epidermidis, which are acne-causing bacteria. In addition, AKE reduced the LPS-induced secretion of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), IL-6, NO and PGE(2) in RAW 264.7 cells, indicating that it has anti-inflammatory effects. Therefore, we suggest that AKE may be an attractive candidate for promoting skin health.
76 citations
TL;DR: Two straightforward methods to detect TAG production in baker’s yeast Saccharomyces cerevisiae are described and demonstrate that a quadruple knockout yeast strain deficient in storage lipids has a reduced growth rate in a medium supplemented with fatty acids.
Abstract: Standard methods to quantify the activity of triacylglycerol (TAG) synthesizing enzymes DGAT and PDAT (TAG-SE) require a sensitive but rather arduous laboratory assay based on radio-labeled substrates. Here we describe two straightforward methods to detect TAG production in baker’s yeast Saccharomyces cerevisiae. First we demonstrate that a quadruple knockout yeast strain deficient in storage lipids has a reduced growth rate in a medium supplemented with fatty acids. This phenotype is rescued by restoring TAG biosynthesis and can be thus used to select yeast cells expressing a recombinant TAG-SE. In the second method, the activity of the recombinant enzyme is measured in a fluorescent in situ assay using Nile red dye that is specific for neutral lipids. Correlation between Nile red fluorescence and enzyme activity is demonstrated with several mutants of a TAG synthesizing enzyme. This yeast live-cell-based assay is rapid, inexpensive, sensitive, and is amenable to high-throughput applications. The methods can be used for a variety of applications such as isolation of novel genes, directed evolution, gene-specific drug screening and will facilitate novel approaches in the research of TAG-SE.
72 citations
TL;DR: IMS system based on MALDI hybrid quadrupole time-of-flight TOF MS revealed the distribution of LysoPtdCho and, more importantly, the organ-specific distribution of TAGs in the embryonic stages of mammals for the first time.
Abstract: Imaging mass spectrometry (IMS) has been developed as a method for determining and visualizing the distribution of proteins and lipids across sections of dissected tissue. Although lipids play an important role in mammal development, their detailed distributions have not been analyzed by conventional methods. In this study, we tried to determine and visualize lysophosphatidylcholine (LysoPtdCho) and triacylglycerol (TAG) in a mouse embryo by matrix-assisted laser desorption/ionization (MALDI) hybrid quadrupole time-of-flight (TOF) mass spectrometer. Many peaks were detected from a raster scan of the whole embryonic sections. The peaks at m/z 496.33, 524.36, 879.72, 881.74, and 921.74 were identified by MS/MS analyses as [LysoPtdCho (16:0) + H]+, [LysoPtdCho (18:0) + H]+, [TAG (16:0/18:2/18:1) + Na]+, [TAG (16:0/18:1/18:1) + Na]+, and [TAG (16:0/20:3/18:1) + K]+, respectively. The ion images constructed from the peaks revealed that LysoPtdCho were distributed throughout the body and TAGs were distributed around the brown adipose tissue and in the liver at embryo day 17.5. Thus, IMS system based on MALDI hybrid quadrupole TOF MS revealed the distribution of LysoPtdCho and, more importantly, the organ-specific distribution of TAGs in the embryonic stages of mammals for the first time. We can conclude that this technique enables us to analyze the roles of various lipids during embryogenesis and gives insight for lipid research.
TL;DR: It is concluded that MetS, in addition to the decrease in HDL-C concentration, is associated with alterations in the HDL phenotype, which is comprised of a greater percentage of small HDL subclasses, which may contribute to the defective antiatherogenic activity of HDL.
Abstract: Patients with metabolic syndrome (MetS) usually have low high density lipoprotein cholesterol (HDL-C) levels. We determined the HDL distribution profile as well as the HDL-related lipoprotein associated phospholipase A2 (HDL-LpPLA2) and paraoxonase-1 (PON1) activities in subjects with MetS (n = 189) but otherwise healthy. Age and sex-matched individuals (n = 166) without MetS served as controls. The lower HDL-C concentration in MetS patients was due to a reduction in both large and small HDL subclasses (P < 0.001 and P < 0.05, respectively). As the number of MetS components increased, the HDL phenotype comprised of a greater percentage of small HDL-3 and less large HDL-2 subclasses, resulting in a decreased HDL-2/HDL-3 ratio (P < 0.001 for all trends). Multivariate analysis revealed that HDL-2 levels and the HDL-2/HDL-3 ratio significantly and independently correlated with HDL-C (positively) and TG (negatively) levels. HDL-3 concentration significantly and independently positively correlated with HDL-C and TG levels. HDL-LpPLA2 activity was decreased in MetS patients (P < 0.01), a phenomenon that may contribute to the defective antiatherogenic activity of HDL in MetS. PON1 activity did not differ between groups. We conclude that MetS, in addition to the decrease in HDL-C concentration, is associated with alterations in the HDL phenotype, which is comprised of a greater percentage of small HDL subclasses. Furthermore, HDL-LpPLA2 activity is decreased in MetS patients.
TL;DR: Linseed supplementation increased the duodenal flow of unsaturated intermediates of biohydrogenation, and this effect was more pronounced for extruded seeds and oil than for rolled seeds, and intestinal digestibility was slightly higher for C and LO diets than for RL and EL.
Abstract: Linseed, a source of linolenic acid, is used in ruminant diets to increase polyunsaturated fatty acids (FA) in animal products. Seed processing is known to have an impact on FA rumen metabolism, but few data are available for linseed. We studied the effect of linseed lipid on ruminal metabolism and intestinal digestibility in cows. Three modes of linseed processing: rolled linseed (RL), extruded linseed (EL) and linseed oil plus linseed meal (LO), supplemented at 7.5% of DM intake, were compared to a control diet (C). Duodenal flows, intestinal digestibility and plasma composition were determined. The duodenal flow of linolenic acid was similar among diets. The sum of t10 and t11-18:1, which were coeluted, was increased with lipid-supplemented diets and represented more than 60% of trans 18:1 for EL and LO diets. The main 18:2 isomers were c9, c12 and t11, c15 among the non-conjugated isomers, and t11, t13 among CLA. Linseed supplementation increased the duodenal flow of unsaturated intermediates of biohydrogenation, and this effect was more pronounced for extruded seeds and oil than for rolled seeds. For most 18-carbon FA, intestinal digestibility was slightly higher for C and LO diets than for RL and EL. Plasma concentrations of non-conjugated 18:2 and linolenic acid were similar among the lipid-supplemented diets. Within diet, profiles of 18:1 isomers (except c9) remained very similar between duodenal and plasma FA.
TL;DR: Octacosadienol was as effective as policosanol in inhibiting the upregulation of HMG coA reductase, in inducing the phosphorylation of AMPK and in downregulating the HMGCoA reduCTase mRNA.
Abstract: Octacosa-10,19-dien-1-ol is a newly synthesized long-chain alcohol, an unsaturated analogue of 1-octacosanol, the major component of policosanol, the purified natural mixture of different higher aliphatic alcohols obtained from sugarcane wax. Our efficient synthetic protocol (five steps with 50% overall yield) is well suited for gram scale preparations and a rapid generation of analogues with different degrees of unsaturation. Beneficial effects of policosanol in the prevention of atherosclerosis and thromboembolic disorders have been reported and related to the inhibition of sterol biosynthesis possibly by the regulation of the activity of HMGCoA reductase mediated by AMP-dependent kinase AMPK. We have compared the effect of octacosadienol and policosanol on the regulation of HMGCoA reductase in HUVEC and HepG2 human hepatoma cells. Octacosadienol was as effective as policosanol in inhibiting the upregulation of HMGCoA reductase, in inducing the phosphorylation of AMPK and in downregulating the HMGCoA reductase mRNA.
TL;DR: An overview of the biological activity of the different classes of PP is given and their analytical applications and the strategies developed so far for the total synthesis of PP are discussed, depending on the synthetic approaches according to the targets and which key steps serve to access the natural products.
Abstract: In animals and plants, fatty acids with at least three double bonds can be oxidized to prostaglandin-like compounds via enzymatic and non-enzymatic pathways. The most common fatty acid precursor in mammals is arachidonic acid (C20:4) (AA) which can be converted through the cyclooxygenase pathway to a series of prostaglandins (PG). Non-enzymatic cyclization of arachidonate yields a series of isoprostanes (IsoP) which comprises all PG (minor compounds) as well as PG isomers that cannot be formed enzymatically. In contrast, in plants, alpha-linolenic acid (C18:3) (ALA) is the most common substrate for the allene oxide synthase pathway leading to the jasmonate (JA) family of lipid mediators. Non-enzymatic oxidation of linolenate leads to a series of C18-IsoPs termed dinor IsoP or phytoprostanes (PP). PP structurally resemble JA but cannot be formed enzymatically. We will give an overview of the biological activity of the different classes of PP and also discuss their analytical applications and the strategies developed so far for the total synthesis of PP, depending on the synthetic approaches according to the targets and which key steps serve to access the natural products.
TL;DR: This study assessed the ability of six strains of bifidobacteria to grow in the presence of α-linolenic acid and to generate conjugated isomers of the fatty acid substrate during fermentation for 42 h and the inhibitory effect of the fermented oils produced.
Abstract: In this study, we assessed the ability of six strains of bifidobacteria (previously shown by us to possess the ability to convert linoleic acid to c9, t11-conjugated linoleic acid (CLA) to grow in the presence of α-linolenic acid and to generate conjugated isomers of the fatty acid substrate during fermentation for 42 h. The six strains of bifidobacteria were grown in modified MRS (mMRS) containing α-linolenic acid for 42 h at 37 °C, after which the fatty acid composition of the growth medium was assessed by gas liquid chromatography (GLC). Indeed, following fermentation of one of the strains, namely Bifidobacterium breve NCIMB 702258, in the presence of 0.41 mg/ml α-linolenic acid, 79.1% was converted to the conjugated isomer, C18:3 c9, t11, c15 conjugated α-linolenic acid (CALA). To examine the inhibitory effect of the fermented oils produced, SW480 colon cancer cells were cultured in the presence of the extracted fermented oil (10–50 μg/ml) for 5 days. The data indicate an inhibitory effect on cell growth (p ≤ 0.001) of CALA, with cell numbers reduced by 85% at a concentration of 180 μM, compared with a reduction of only 50% with α-linolenic acid (p ≤ 0.01).
TL;DR: A fast, efficient, and sensitive liquid chromatography–mass spectrometry method to quantify eight different classes of gangliosides in the brains of 2- day-old and 80-day-old Wistar rats, showing good repeatability and mean recoveries.
Abstract: Gangliosides are a large family of glycosphingolipids that are abundant in the brain, and have been shown to affect neuronal plasticity during development, adulthood, and aging. We developed a fast, efficient, and sensitive liquid chromatography–mass spectrometry method to quantify eight different classes of gangliosides (GM1, GM2, GM3, GD3, GD1a, GD1b, GT1b, GQ1b) in the brains of 2-day-old and 80-day-old Wistar rats. The gangliosides were extracted from rat brain using a modified Svennerholm and Fredman method. After ganglioside class separation using a hydrophilic high performance liquid chromatography (HPLC) column, the resolving power of the LTQ-Orbitrap™ mass spectrometer was used to extract and sum the major species of each ganglioside class, generating fully resolved extracted ion current peaks for both standards and samples. The flexibility and the specificity of this method are such that it can be applied to the analysis of other ganglioside species/classes not discussed in this paper, provided appropriate standards are available. The method had good repeatability (coefficient of variation 4.8–12.3%) and mean recoveries in the range 92–107%.
TL;DR: A practical method to quantify the simultaneous transfer to HDL of phospholipids, free-cholesterol, esterified cholesterol and triacylglycerols and to verify the lipid transfer in patients with coronary artery disease or undergoing statin treatment was tested.
Abstract: The exchange of lipids with cells and other lipoproteins is a crucial process in HDL metabolism and for HDL antiatherogenic function. Here, we tested a practical method to quantify the simultaneous transfer to HDL of phospholipids, free-cholesterol, esterified cholesterol and triacylglycerols and to verify the lipid transfer in patients with coronary artery disease (CAD) or undergoing statin treatment. Twenty-eight control subjects without CAD, 27 with CAD and 25 CAD patients under simvastatin treatment were studied. Plasma samples were incubated with a donor nanoemulsion prepared by ultrasonication of the constituent lipids and labeled with radioactive lipids; % lipids transferred to HDL were quantified in the HDL-containing supernatant after chemical precipitation of non-HDL fractions and the nanoemulsion. The assay was precise and reproducible. Increase of temperature (4–37 °C), of incubation period (5 min to 2 h), of HDL-cholesterol concentration (33–244 mg/dL) and of mass of nanoemulsion lipids (0.075–0.3 mg/μL) resulted in increased lipid transfer from the nanoemulsion to HDL. In contrast, increasing pH (6.5–8.5) and albumin concentration (3.5–7.0 g/dL) did not affect lipid transfer. There was no difference between CAD and control non-CAD with regard to the lipid transfer, but statin treatment reduced the transfer to HDL of all four lipids. The test herein described is a valid and practical tool for exploring an important aspect of HDL metabolism.
TL;DR: By limiting exogenous lipid availability and inhibiting FAS using Orlistat, this work demonstrated both a greater sensitivity and amplified cancer cell death by activation of apoptosis and identified “windows of opportunity” at which time apoptosis can be aborted and cells can be reversed from the death pathway.
Abstract: Orlistat, an anti-obesity drug, is a potent inhibitor of fatty acid synthase (FAS) and tumor cell viability. It can also induce apoptotic cancer cell death. We examined the effects of Orlistat on cultured NUGC-3 gastric cancer cells. We identified that inhibition of FAS via Orlistat exposure results in rapid cellular damage preceded by a direct but short-lived autophagic response. The Orlistat induced damage can be reversed through the addition of lipid containing media in a process that normally leads to cell death. By limiting exogenous lipid availability and inhibiting FAS using Orlistat, we demonstrated both a greater sensitivity and amplified cancer cell death by activation of apoptosis. We have identified "windows of opportunity" at which time apoptosis can be aborted and cells can be reversed from the death pathway. However, when challenged beyond the window of recovery, cell death becomes all but certain as the ability to be rescued decreases considerably. In vivo examination of Orlistat's ability to inhibit gastrointestinal cancer was examined using heterozygous male C57BL/6J APC-Min mice, which spontaneously develop a fatal gastrointestinal cancer. Mice were fed either a high fat (11%) or low fat (1.2%) diet containing no Orlistat or 0.5 mg Orlistat/g of chow. Orlistat treated mice fed the high fat, but not low fat diet, survived 7-10% longer than the untreated controls.
TL;DR: It is hypothesize that the effect of plant stanol esters on serum TAG concentrations origins from a lowered hepatic production of large TAG-rich VLDL-1 particles.
Abstract: Plant stanol esters not only lower low density lipoprotein cholesterol but also have previously been shown to lower serum triacylglycerol (TAG) concentrations, especially in subjects with elevated TAG concentrations. To find a possible explanation, we explored changes in serum lipoprotein profiles, as measured with nuclear magnetic resonance. For this, serum samples from two parallel-designed controlled studies were evaluated before and 8 weeks after the consumption of plant stanol esters. In the first study, dyslipidemic metabolic syndrome subjects participated and in the second study normolipidemic subjects. In metabolic syndrome subjects, plant stanol esters lowered concentrations of large (>60 nm) and medium (35-60 nm) VLDL particles as compared to controls. In normolipidemic subjects, the serum concentration of large VLDL-1 particles was also lowered, although less pronounced. Based on these findings, we hypothesize that the effect of plant stanol esters on serum TAG concentrations origins from a lowered hepatic production of large TAG-rich VLDL-1 particles.
TL;DR: Markers of inflammation were related to plaque size alone, implying inflammation to be predominantly associated with the amount of atherosclerosis, and suggest that plaque size and echogenicity are influenced by different risk factors.
Abstract: Carotid plaques can be characterised by ultrasound by size and echogenicity. Both size and echogenicity are predictors of cardiovascular events. The aim of this study was to examine whether traditional risk factors and markers of inflammation and oxidation were associated with plaque size and echogenicity. Computerised analysis of carotid plaque size and echogenicity (grey scale median, GSM) were performed by ultrasound in a population-based health survey in 1,016 subjects aged 70 years (PIVUS study). Information on cardiovascular risk factors was collected, together with markers of inflammation and oxidation. Increased Framingham risk score, systolic blood pressure, higher BMI and decreased HDL, lower glutathione levels were related to echolucent plaques. Previous or present smoking was common with significantly more pack-years related to the echorich plaques. Plaque size was associated with increased Framingham risk score, systolic blood pressure, blood glucose levels, smoking, ApoB/A1 ratio, OxLDL, TNF alpha, HOMA insulin resistance, leucocyte count, decreased BCD-LDL and low levels of l-selectin. Low HDL, increased BMI and decreased glutathione levels were associated with the echolucency of carotid plaques, implying metabolic factors to play a role for plaque composition. Markers of inflammation were related to plaque size alone, implying inflammation to be predominantly associated with the amount of atherosclerosis. These results suggest that plaque size and echogenicity are influenced by different risk factors.
TL;DR: The levels of fatty acids in human milk may reflect the current diet of the mother as well as the diet consumed early in pregnancy, and margarines, bakery products and confectionery are a major source of trans fatty acid in maternal diet in Turkey.
Abstract: The aim of this study was to determine the fatty acid composition and trans fatty acid and fatty acid contents of breast milk in Turkish women and to find the effect of breastfeeding mothers’ diet on trans fatty acid and fatty acid composition Mature milk samples obtained from 50 Turkish nursing women were analyzed Total milk lipids extracts were transmethylated and analyzed by using gas liquid chromatography to determine fatty acids contents A questionnaire was applied to observe eating habits and 3 days dietary records from mothers were obtained Daily dietary intake of total energy and nutrients were estimated by using nutrient database The mean total trans fatty acids contents was 213 ± 103% The major sources of trans fatty acids in mothers’ diets were margarines-butter (370%), bakery products and confectionery (296%) Mothers who had high level of trans isomers in their milk consumed significantly higher amounts of these products Saturated fatty acids, polyunsaturated fatty acids and monounsaturated fatty acids of human milk constituted 407 ± 47%, 269 ± 42% and 308 ± 06% of the total fatty acids, respectively The levels of fatty acids in human milk may reflect the current diet of the mother as well as the diet consumed early in pregnancy Margarines, bakery products and confectionery are a major source of trans fatty acids in maternal diet in Turkey
TL;DR: This study is the first comparative analysis of brain lipids in mice, cats, and humans with SD and will be important for designing therapies for Sandhoff disease patients.
Abstract: Sandhoff disease (SD) is a glycosphingolipid (GSL) storage disease that arises from an autosomal recessive mutation in the gene for the β-subunit of β-Hexosaminidase A (Hexb gene), which catabolizes ganglioside GM2 within lysosomes Accumulation of GM2 and asialo-GM2 (GA2) occurs primarily in the CNS, leading to neurodegeneration and brain dysfunction We analyzed the total lipids in the brains of SD mice, cats, and humans GM2 and GA2 were mostly undetectable in the normal mouse, cat, and human brain The lipid abnormalities in the SD cat brain were generally intermediate to those observed in the SD mouse and the SD human brains GM2 comprised 38, 67, and 87% of the total brain ganglioside distribution in the SD mice, cats, and humans, respectively The ratio of GA2–GM2 was 093, 013, and 027 in the SD mice, cats, and humans, respectively, suggesting that the relative storage of GA2 is greater in the SD mouse than in the SD cat or human Finally, the myelin-enriched lipids, cerebrosides and sulfatides, were significantly lower in the SD brains than in the control brains This study is the first comparative analysis of brain lipids in mice, cats, and humans with SD and will be important for designing therapies for Sandhoff disease patients
TL;DR: The wild type SLC1 was expressed in soybean somatic embryos under the control of seed specific phaseolin promoter and Expression of the yeast Wt LPAT gene did not alter the fatty acid composition of the seed oil.
Abstract: The wild type (Wt) and mutant form of yeast (sphingolipid compensation) genes, SLC1 and SLC1-1, have been shown to have lysophosphatidic acid acyltransferase (LPAT) activities (Nageic et al. in J Biol Chem 269:22156-22163, 1993). Expression of these LPAT genes was reported to increase oil content in transgenic Arabidopsis and Brassica napus. It is of interest to determine if the TAG content increase would also be seen in soybeans. Therefore, the wild type SLC1 was expressed in soybean somatic embryos under the control of seed specific phaseolin promoter. Some transgenic somatic embryos and in both T2 and T3 transgenic seeds showed higher oil contents. Compared to controls, the average increase in triglyceride values went up by 1.5% in transgenic somatic embryos. A maximum of 3.2% increase in seed oil content was observed in a T3 line. Expression of the yeast Wt LPAT gene did not alter the fatty acid composition of the seed oil.
TL;DR: It is indicated that seal oil is as effective as fish oil in lowering plasma triglycerides and blood pressure.
Abstract: As meat is a rich source of the omega-3 fatty acid docosapentaenoic acid (DPA) and Australians consume six times more meat than fish, investigation of the potential health benefit of DPA is warranted. The aims were to compare the effects of seal oil supplementation with fish oil, on measures of plasma lipids and blood pressure in hypertriglyceridaemic subjects. Forty-eight volunteers were recruited from the Wollongong community and were randomly allocated to one of three groups either receiving 1 g/day of long-chain omega-3 polyunsaturated fatty acids (LC n-3 PUFA) using one of three oils: seal oil capsules (340 mg eicosapentaenoic acid (EPA), 230 mg DPA, 450 mg DHA), fish oil capsules (210 mg EPA, 30 mg DPA, 810 mg DHA) or placebo capsules (containing sunola oil) for 6 weeks. Plasma triglycerides remained unchanged in the placebo group, whilst reductions of 7 and 14% (P < 0.05) were seen in the fish oil and seal oil groups respectively. Systolic blood pressure improved by 8 and 5 mmHg with seal oil and fish oil respectively (P < 0.05). The mean arterial pressure was significantly lower after seal oil supplementation (P < 0.005) compared with the placebo group. These results indicate that seal oil is as effective as fish oil in lowering plasma triglycerides and blood pressure.
TL;DR: It is demonstrated that the t10,c12 CLA isomer inhibits growth of MCF-7 cells and suggested that this may be mediated through incorporation into cellular phospholipids and interference with the function of IGF-I and related signaling proteins.
Abstract: In vitro work suggests that conjugated linoleic acid (CLA) isomers (c9,t11 and t10,c12) are cytotoxic to human breast cancer cells, however the mechanism remains unknown. Using human MCF-7 breast cancer cells, we examined the effects of c9,t11 and t10,c12 CLA compared to oleic acid (OA), linoleic acid (LA), or untreated cells on cell membrane phospholipid composition, cell survival, and the insulin-like growth factor-I (IGF-I) and the downstream insulin receptor substrate-1 (IRS-1). Both CLA isomers were incorporated into membrane phospholipids (p < 0.05). Compared to untreated cells, c9,t11 or t10,c12 CLA significantly reduced the metabolic activity of IGF-I stimulated MCF-7 cells, increased lactate dehydrogenase (LDH) release, and decreased cellular concentrations of the IGF-I receptor (IGF-IR) and insulin receptor substrate-1 (p < 0.05). Incubation with t10,c12 CLA also reduced the levels of phosphorylated IGF-1R. The effects on all of these measures were greater (p < 0.05) for t10,c12 CLA compared to c9,t11 CLA. There were few differences between LA-treated and c9,t11 CLA-treated cells, whereas cellular metabolic activity, LDH release, and IGF-IR concentrations differed between t10,c12 CLA-treated and LA-treated cells (p < 0.05). OA stimulated growth compared to the untreated condition (p < 0.05). In summary, this study demonstrated that the t10,c12 CLA isomer inhibits growth of MCF-7 cells and suggested that this may be mediated through incorporation into cellular phospholipids and interference with the function of IGF-I and related signaling proteins.
TL;DR: Data show that enhancing n-3 PUFA enrichment in muscle leads to significant increase in IMF content, which may be due to alterations in the expression of genes involved in adipogenesis, however this will need to be confirmed by protein and enzyme activity studies.
Abstract: The aim of the study was to investigate the effect of n-3 PUFA enrichment in longissimus muscle on intramuscular fat (IMF) content and expression of related genes in growing-finishing barrows. Two isoenergetic, isonitrogenous and isolipidic diets were formulated: one was basal diet and the other contained 10% linseed. Twenty-four Landrace x NewDamLine barrows weighing 35 +/- 3.7 kg were randomly assigned to four treatment groups with six pigs per group. During the whole experimental period of 90 days, all groups were first fed the basal diet and then the linseed diet for 0, 30, 60, and 90 days before slaughter, respectively. Meat quality, fatty acid composition, and expression of genes involved in adipogenesis in longissimus muscle were measured and analyzed. The IMF content increased linearly (P < 0.05) as the linseed diet feeding time prolonged. Meanwhile, n-3 PUFA content and expression of peroxisome proliferator-activated receptor delta (PPARdelta), PPARgamma, adipocyte fatty acid-binding protein (aP2) and lipoprotein lipase (LPL) increased linearly (P < 0.01) as well, while the expression of wingless related MMTV integration site 10b (Wnt10b) linearly decreased (P < 0.01). Furthermore, significant (P < 0.01) quadratic or linear relation was observed between n-3 PUFA enrichment and expression of these genes, while significant (P < 0.01) quadratic or linear relation was observed between the expression of PPARgamma, aP2 or Wnt10b and IMF content. These data show that enhancing n-3 PUFA enrichment in muscle leads to significant increase in IMF content. A possible explanation is due to alterations in the expression of genes involved in adipogenesis, however this will need to be confirmed by protein and enzyme activity studies.
TL;DR: The levels of undesirable TFA and CLA isomers in human milk are a concern and efforts to decrease the practice of high temperature stir-frying using unsaturated oils, and a promotion to increase consumption of dairy and ruminant products should be considered in China.
Abstract: Human milk was obtained from 97 healthy lactating women from five different regions in China. Twenty-four hour dietary questionnaire identified the foods consumed that showed distinct differences in food types between cities. The southern and central regions had higher levels of total trans fatty acids (TFA) and conjugated linoleic acids (CLA) in human milk than the northern region. The major isomers in human milk from the northern region were vaccenic and rumenic acids, whereas the other regions had a random distribution of these isomers. This was consistent with the isomer distribution in the refined vegetable oils used and their increased formation during high temperature stir-frying. The human milk composition showed little evidence that partially hydrogenated fats were consumed, except eight mothers in Guangzhou who reported eating crackers, plus four other mothers. The TFA concentration in these human milk samples was higher (2.06-3.96%). The amount of n-6 (1.70-2.24%) and n-3 (0.60-1.47%) highly unsaturated fatty acids (HUFA) in human milk and the resultant ratio (1.43-2.95) showed all mothers in China had an adequate supply of HUFA in their diet. Rapeseed oil was consumed evidenced by erucic acids in human milk. The levels of erucic acid were below internationally accepted limits for human consumption. The levels of undesirable TFA and CLA isomers in human milk are a concern. Efforts to decrease the practice of high temperature stir-frying using unsaturated oils, and a promotion to increase consumption of dairy and ruminant products should be considered in China.
TL;DR: GC analysis for determining the origin of North American salmon compared favorably with the astaxanthin isomer technique used by the FDA and showed that the fatty acid 18:2n-6 was the key indicator associated with theorigin of these salmon.
Abstract: Mislabeling of farmed and wild salmon sold in markets has been reported. Since the fatty acid content of fish may influence human health and thus consumer behavior, a simplified method to identify wild and farmed salmon is necessary. Several studies have demonstrated differences in lipid profiles between farmed and wild salmon but no data exists validating these differences with government-approved methods to accurately identify the origin of these fish. Current methods are both expensive and complicated, using highly specialized equipment not commonly available. Therefore, we developed a testing protocol using gas chromatography (GC), to determine the origin of salmon using fatty acid profiles. We also compared the GC method with the currently approved FDA (United States Food and Drug Administration) technique that uses analysis of carotenoid optical isomers and found 100% agreement. Statistical validation (n = 30) was obtained showing elevated 18:2n-6 (z = 4.56; P = 0.0001) and decreased 20:1n-9 (z = 1.79; P = 0.07) in farmed samples. The method is suitable for wide adaptation because fatty acid methyl ester analysis is a well-established procedure in labs that conduct analysis of lipid composition and food constituents. GC analysis for determining the origin of North American salmon compared favorably with the astaxanthin isomer technique used by the FDA and showed that the fatty acid 18:2n-6 was the key indicator associated with the origin of these salmon.
TL;DR: It is demonstrated that the synergistic antiproliferative effects of combined low dose statin and γ-tocotrienol treatment are directly related to an inhibition in HMGR activity and subsequent suppression in mevalonate synthesis.
Abstract: Statins directly inhibit 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR) activity, while γ-tocotrienol, an isoform of vitamin E, enhances the degradation and reduces cellular levels of HMGR in various tumor cell lines. Since treatment with statins or γ-tocotrienol alone induced a dose-responsive inhibition, whereas combined treatment with subeffective doses of these agents resulted in a synergistic inhibition in +SA mammary tumor cell growth, studies were conducted to investigate the role of the HMGR pathway in mediating the antiproliferative effects of combined low dose statin and γ-tocotrienol. Treatment with 8 μM simvastatin inhibited cell growth and isoprenylation of Rap1A and Rab6, and supplementation with 2 μM mevalonate reversed these effects. However, the growth inhibitory effects of 4 μM γ-tocotrienol were not dependent upon suppression in mevalonate synthesis. Treatment with subeffective doses of simvastatin (0.25 μM), lovastatin (0.25 μM), mevastatin (0.25 μM), pravastatin (10 μM), or γ-tocotrienol (2 μM) alone had no effect on protein prenylation or mitogenic signaling, whereas combined treatment with these agents resulted in a significant inhibition in +SA cell growth, and a corresponding decrease in total HMGR, Rap1A and Rab6 prenylation, and MAPK signaling, and mevalonate supplementation reversed these effects. These findings demonstrate that the synergistic antiproliferative effects of combined low dose statin and γ-tocotrienol treatment are directly related to an inhibition in HMGR activity and subsequent suppression in mevalonate synthesis.