bluuebunny/crossref_metadata_2025 huggingface dataset:
Distribution of medium‐chain FA in different lipid classes after administration of specific structured TAG in rats
Authors: Mu Huiling , Høy Carl‐Erik
Published: November 2006

Structured TAG (STAG) containing medium‐chain FA (MCFA) in the sn‐1,3 positions and essential FA in the sn‐2 position were synthesized by lipase‐catalyzed acidolysis. In our previous studies we found that part of the MCFA from STAG could be absorbed in the small intestine; however, it was unclear how they were absorbed. In order to get a better understanding of the metabolism of STAG to improve future design and application of STAG, in the present study lymph lipids collected after feeding STAG were fractionated into different classes and the FA composition of each lipid class was studied by GC after methylation to FAME. Caprylic acid was detected in the fraction of TAG only after administration of 1,3‐dioctanoyl‐2‐linoleyl‐sn‐glycerol (8∶0/18∶2/8∶0), whereas lauric acid was detected in TAG, DAG, and FFA as well as phospholipids after administration of 1,3‐didodecanoyl‐2‐linoleyl‐sn‐glycerol (12∶0/18∶2/12∶0). We conclude that the enterocyte has the ability to reacylate the MCFA into TAG and that the intestinal absorption of MCFA from STAG mainly occurs by resynthesis of TAG. Caprylic acid from STAG is not incorporated into phospholipids, whereas lauric acid from STAG can be incorporated into phospholipids.
Habitual fish intake is associated with decreased LDL susceptibility to ex vivo oxidation
Authors: de Gordoa Juan Carlos Ruiz , de Renobales Mertxe , del Cerro Ana , de Labastida Eva Fernández , Amiano Pilar , Dorronsoro Miren ,   the EPIC Group of Spain E
Published: November 2006

A sample of 101 free‐living individuals eating their habitual diets had fish consumptions ranging from less than one serving per week to over five servings per week. Statistically significant positive correlations were found between the amounts of EPA (20:5), DHA (22:6), and total n‐3 PUFA ingested with the diet and their amounts in serum and in the phospholipid and cholesterol ester fractions of isolated LDL. No statistically significant correlations were observed between the intake and the serum or LDL amounts of any other FA [total n‐6 PUFA, linoleic acid (18:2), arachidonic acid (20:4), monounsaturated FA, or saturated FA)]. The increase in serum n‐3 PUFA did not affect the Trolox‐equivalent antioxidant capacity of serum (1.18 ±0.17 mmol/l). When isolated LDI were subjected to Cu2+‐induced ex vivo oxidation, a statistically significant but negative correlation was found between intake of n‐3 PUFA and the rate of appearance of conjugated dienes as well as with the total amount of conjugated dienes. In contrast, intake of n‐6 PUFA showed a significant and positive correlation with these two oxidation parameters. The observed results suggest that 22:6 but not 20:5 could have a possible protective effect, whereas perhaps 20:4 and 18:2 could have a prooxidant effect.
Dietary conjugated linolenic acid in relation to CLA differently modifies body fat mass and serum and liver lipid levels in rats
Authors: Koba Kazunori , Akahoshi Asuka , Yamasaki Masao , Tanaka Kazunari , Yamada Koji , Iwata Toshio , Kamegai Takeshi , Tsutsumi Kentaro , Sugano Michihiro
Published: November 2006

The present study compared the effect of dietary conjugated linolenic acid (CLNA) on body fat and serum and liver lipid levels with that of CLA in rats. FFA rich in linoleic acid, α‐linolenic acid, CLA, or CLNA were used as experimental fats. Male Sprague‐Dawley rats (4 wk old) were fed purified diets containing 1% of one of these experimental fats. After 4 wk of feeding, adipose tissue weights, serum and liver lipid concentrations, serum tumor necrosis factor (TNF)‐α and leptin levels, and hepatic β‐oxidation activities were measured. Compared with linoleic acid, CLA and, more potently, CLNA were found to reduce perirenal adipose tissue weight. The same trend was observed in the weight of epididymal adipose tissue. CLNA, but not CLA, was found to significantly increase serum and liver IG concentrations. Serum FFA concentration was also increased in the CLNA group more than in the other groups. The activity of β‐oxidation in liver mitochondria and peroxisomes was significantly higher in the CLNA group than in the other groups. Thus, the amount of liver TG exceeded the ability of hepatic β‐oxidation. Significant positive correlation was found between the adipose tissue weights and serum leptin levels in all animals (vs. perirenal: r=0.557, P<0.001; vs. epididymal: r=0.405, P<0.05). A less significant correlation was found between adipose tissue weights and serum TNF‐α level (vs. perirenal: r=0.069, P<0.1; vs. epididymal: r=0.382, P<0.05). Although the mechanism for the specific effect of CLNA is not clear at present, these findings indicate that in rats CLNA modulated the body fat and TG metabolism differently from CLA.
Dose effect of α‐linolenic acid on PUFA conversion, bioavailability, and storage in the hamster
Authors: Morise Anne , Combe Nicole , Boué Carole , Legrand Philippe , Catheline Daniel , Delplanque Bernadette , Fénart Evelyne , Weill Pierre , Hermier Dominique
Published: November 2006

If an increased consumption of α‐linolenic acid (ALA) is to be promoted in parallel with that of n−3 long‐chain‐rich food, it is necessary to consider to what extent dietary ALA can be absorbed, transported, stored, and converted into long‐chain derivatives. We investigated these processes in male hamsters, over a broad range of supply as linseed oil (0.37, 3.5, 6.9, and 14.6% energy). Linoleic acid (LA) was kept constant (8.5% energy), and the LA/ALA ratio was varied from 22.5 to 0.6. The apparent absorption of individual FA was very high (>96%), and that of ALA remained almost maximum even at the largest supply (99.5%). The capacity for ALA transport and storage had no limitation over the chosen range of dietary intake. Indeed, ALA intake was significantly correlated with ALA level not only in cholesteryl esters (from 0.3 to 9.7% of total FA) but also in plasma phospholipids and red blood cells (RBC), which makes blood components extremely reliable as biomarkers of ALA consumption. Similarly, ALA storage in adipose tissue increased from 0.85 to 14% of total FA and was highly correlated with ALA intake. As for bioconversion, dietary ALA failed to increase 22∶6n−3, decreased 20∶4n−6, and efficiently increased 20∶5n−3 (EPA) in RBC and cardiomyocytes. EPA accumulation did not tend to plateau, in accordance with identical activities of Δ5‐ and Δ6‐desaturases in all groups. Dietary supply of ALA was therefore a very efficient means of improving the 20∶4n−6 to 20∶5n−3 balance.
A critical evaluation of Raman spectroscopy for the analysis of lipids: Fatty acid methyl esters
Authors: Beattie J. Renwick , Bell Steven E. J. , Moss Bruce W.
Published: November 2006

The work presented here is aimed at determining the potential and limitations of Raman spectroscopy for fat analysis by carrying out a systematic investigation of C4−C24 FAME. These provide a simple, well‐characterized set of compounds in which the effect of making incremental changes can be studied over a wide range of chain lengths and degrees of unsaturation. The effect of temperature on the spectra was investigated over much larger ranges than would normally be encountered in real analytical measurements. It was found that for liquid FAME the best internal standard band was the carbonyl stretching vibration ρ(C=O), whose position is affected by changes in sample chain length and physical state; in the samples studied here, it was found to lie between 1729 and 1748 cm−1. Further, molar unsaturation could be correlated with the ratio of the ρ(C=O) to either ρ(C=C) or δ(H−C=) with R2>0.995. Chain length was correlated with the δ(CH2)tw/ρ(C=O) ratio, (where “tw” indicates twisting) but separate plots for odd‐ and even‐numbered carbon chains were necessary to obtain R2>0.99 for liquid samples. Combining the odd‐ and even‐numbered carbon chain data in a single plot reduced the correlation to R2=0.94–0.96, depending on the band ratios used. For molal unsaturation the band ratio that correlated linearly with unsaturation (R2>0.99) was ρ(C=C)/δ(CH2)sc (where “sc” indicates scissoring). Other band ratios show much more complex behavior with changes in chemical and physical structure. This complex behavior results from the fact that the bands do not arise from simple vibrations of small, discrete regions of the molecules but are due to complex motions of large sections of the FAME so that making incremental changes in structure does not necessarily lead to simple incremental changes in spectra.
Dietary intake of essential and long‐chain polyunsaturated fatty acids in pregnancy
Authors: Loosemore Elizabeth D. , Judge Michelle P. , Lammi‐Keefe Carol J.
Published: November 2006

The dietary intake of EFA and long‐chain PUFA (LCPUFA) by women with (n=14) and without (n=31) gestational diabetes mellitus (GDM) was determined by repeated 24‐h recalls. Women with GDM consumed significantly more energy as fat compared with women who had uncomplicated pregnancies; absolute dietary fat did not differ. Dietary n−3 LCPUFA was substantially lower than the current recommendation for pregnancy, whereas intake of saturated FA (SFA) exceeded it. We conclude that replacing dietary sources of SFA with those of EFA and LCPUFA, especially n−3 LCPUFA, would benefit the dietary fat profiles of all pregnant women.
Low concentration of oxidized low density lipoprotein suppresses platelet reactivity in vitro: An intracellular study
Authors: Chou Duen‐Suey , Hsiao George , Shen Ming‐Yi , Fong Tsorng‐Harn , Lin Chien‐Huang , Chen Tzeng‐Fu , Sheu Joen‐Rong
Published: November 2006

The intracellular mechanisms underlying oxidized low density lipoprotein (oxLDL)‐signaling pathways in platelets remain obscure and findings have been controversial. Therefore, we examined the influence of oxLDL in washed human platelets. In this study oxLDL concentration‐dependently (20–100 μg/mL) inhibited platelet aggregation in human platelets stimulated by collagen (1 μg/mL) and arachidonic acid (60 μM), but not by thrombin (0.02 U/mL). The activity of oxLDL was greater at 24 h in inhibiting platelet aggregation than at 12 h. At 24 h, oxLDL concentration‐dependently inhibited intracellular Ca2+ mobilization and thromboxane B2 formation in human platelets stimulated by collagen. In addition, at 24 h oxLDL (40 and 80 μg/mL) significantly increased the formation of cyclic AMP, but not cyclic GMP or nitrate. In an ESR study, 24 h‐oxLDL (40 μg/mL) markedly reduced the ESR signal intensity of hydroxyl radicals (OH−) in both collagen (2 μg/mL)‐activated platelets and Fenton reaction (H2O2+Fe2+). The inhibitory effect of oxLDL may induce radical‐radical termination reactions by oxLDL‐derived lipid radical interactions with free radicals (such as hydroxyl radicals) released from activated platelets, with a resultant lowering of intracellular Ca2+ mobilization followed by inhibition of thromboxane A2 formation, thereby leading to increased cyclic AMP formation and finally inhibited platelet aggregation. This study provides new insights concerning the effect of oxLDL in platelet aggregation.
Dietary TAG source and level affect performance and lipase expression in larval sea bass (Dicentrarchus labrax)
Authors: Morais S. , Cahu C. , Zambonino‐Infante J. L. , Robin J. , Rønnestad I. , Dinis M. T. , Conceição L. E. C.
Published: November 2006

The influence of dietary TAG source (fish oil, triolein, and coconut oil) and level (7.5 and 15% of the diet) on growth, lipase activity, and mRNA level was studied in sea bass larvae, from mouth opening until day 24 and from day 37 to 52. Fish oil and triolein induced better growth in both experiments, this being significant at a higher dietary level. Coconut oil significantly decreased growth at the higher level, possibly as the result of an excessive supply of medium‐chain TAG. Growth was not related to lipase specific activity, suggesting a production in excess to dietary needs. Body lipid content was positively related to dietary lipid level and was affected by lipid quality. In addition, larval FA composition generally reflected that of the diet. The source of dietary lipid, but not the quantity, was shown to affect lipase activity significantly. Coconut oil diets induced the highest lipase activity, whereas the effect of fish oil was age dependent‐it was similar to coconut oil at day 24 but induced the lowest lipase activity in 52‐d‐old larvae. The differential lipase response was probably caused by differences in the FA composition of the diet, related to the specificity of lipase toward FA differing in chain length and degree of saturation. No significant differences were found in lipase/glyceraldehyde‐3‐phosphate dehydrogenase mRNA, which suggests the existence of a posttranscriptional regulation mechanism.
Distribution of tocopherols and tocotrienols to rat ocular tissues after topical ophthalmic administration
Authors: Tanito Masaki , Itoh Nanako , Yoshida Yasukazu , Hayakawa Mieko , Ohira Akihiro , Niki Etsuo
Published: November 2006

With increasing evidence suggesting the involvement of oxidative stress in various disorders and diseases, the role of antioxidants in vivo has received much attention. Chemically, tocopherols and tocotrienols are closely related; however, it has been observed that they have widely varying degrees of biological effectiveness. The present study has been carried out in an attempt to deepen our understanding of whether there is a significant difference in distribution between tocopherol and tocotrienol homologs to rat eye tissues. Rats were administered 5 μl of pure tocopherol or tocotrienol to each eye once a day for 4 d. Various tissues of the eyes were separated and analyzed for tocopherol and tocotrienol concentrations. The concentration of α‐tocotrienol increased markedly in every tissue to which it was administered; however, no significant increase was observed in the case of α‐tocopherol. The intraocular penetration of γ‐tocopherol and γ‐tocotrienol did not differ significantly. Additionally, a significant increase in total vitamin E concentration was observed in ocular tissues, including crystalline lens, neural retina, and eye cup, with topical administration using a relatively small amount (5 μL) of vitamin E, whereas no significant increase was observed when the same amount of vitamin E was administered orally. Topical administration of tocotrienols is thus an effective way to increase ocular tissue vitamin E concentration.
Formation of genotoxic dicarbonyl compounds in dietary oils upon oxidation
Authors: Fujioka Kazutoshi , Shibamoto Takayuki
Published: November 2006

Dietary oils—tuna, salmon, cod liver, soybean, olive, and corn oils—were treated with accelerated storage conditions (60°C for 3 and 7 d) and a cooking condition (200°C for 1 h). Genotoxic malonaldehyde (MA), glyoxal, and methylglyoxal formed in the oils were analyzed by GC. Salmon oil produced the greatest amount of MA (1070±77.0 ppm of oil) when it was heated at 60°C for 7 d. The highest formation of glyoxal was obtained from salmon oil heated at 60°C for 3 d. More glyoxal was found from salmon and cod liver oils when they were heated for 3 d (12.8±1.10 and 7.07±0.19 ppm, respectively) than for 7d (6.70±0.08 and 5.94±0.38 ppm, respectively), suggesting that glyoxal underwent secondary reactions during a prolonged time. The amount of methyglyoxal formed ranged from 2.03±0.13 (cod liver oil) to 2.89±0.11 ppm (tuna oil) in the fish oils heated at 60°C for 7 d. Among vegetable oils, only olive oil yielded methylglyoxal (0.61±0.03 ppm) under accelerated storage conditions. When oils were treated under cooking conditions, the aldehydes formed were comparable to those formed under accelerated storage conditions. Fish oils produced more MA, glyoxal, and methylglyoxal than did vegetable oils because the fish oils contained higher levels of long‐chain PUFA, such as EPA and DHA, than did the vegetable oils. A statistically significant correlation (P<0.05) between the α‐tocopherol content and the oxidation parameters was obtained from only MA and fish oils heated at 60°C for 3 d.
Fatty acids of the seeds of Origanum onites L. and O. vulgare L.
Authors: Azcan Nezihe , Kara Mustafa , Demirci Betül , Başer K. Hüsnü Can
Published: November 2006

Seed oils of Origanum onites L. from the Antalya and Mugla regions and O. vulgare L. from the Kirklareli region of Turkey were extracted with hexane in a Soxhlet apparatus. The oil yields were 14.1–20.0 and 18.5%, respectively. FA compositions of the seed oils were determined by GC and GC/MS. Twenty FA were identified in both O. onites and O. vulgare seeds. The major FA of both species were linolenic (56.3–57.0%; 61.8%), linoleic (21.5–21.7%; 18.8%), oleic (8.7–8.9%; 5.9%), palmitic (5.9–6.5%; 5.5%), stearic (2.1–2.4%; 2.1%), and (Z)‐11‐octadecenoic (0.6–0.8%; 0.5%), respectively.
New approach to the analysis of oxidized triacylglycerols in lipoproteins
Authors: Suomela Jukka‐Pekka , Ahotupa Markku , Sjövall Olli , Kurvinen Juha‐Pekka , Kallio Heikki
Published: November 2006

The oxidation of human LDL lipids and the structures of oxidized TAG molecules found in LDL were investigated. Pooled samples of 10 normolipidemic and 10 hyperlipidemic subjects were analyzed. For determination of the oxidation levels, the LDL baseline diene conjugation (LDL‐BDC) method was used. A method based on HPLC and electrospray ionization‐MS was optimized and applied to the analysis of molecular structures of oxidized TAG in LDL. Differences were found between the oxidation levels of the samples. The LDL‐BDC value was 22.2 μmol/L serum in the normolipidemic group, and 88.1 μmol/L serum in the hyperlipidemic group. The amounts of oxidized TAG molecules were small. However, several species of oxidized TAG were identified. These included TAG molecules with a keto or an epoxy group attached to a FA, and TAG molecules with a FA core aldehyde. In some TAG, the keto/epoxy ratio was greater in the hyperlipidemic group compared to the normolipidemic group. The results show that our approach is applicable to research on lipid oxidation in lipoproteins.
Protein engineering and applications of Candida rugosa lipase isoforms
Authors: Akoh Casimir C. , Lee Guan‐Chiun , Shaw Jei‐Fu
Published: November 2006

Commercial preparations of Candida rugosa lipase (CRL) are mixtures of lipase isoforms used for the hydrolysis and synthesis of various esters. The presence of variable isoforms and the amount of lipolytic protein in the crude lipase preparations lead to a lack of reproducibility of biocatalytic reactions. Purification of crude CRL improve their substrate specificity, enantioselectivity, stability, and specific activities. The expression of the isoforms is governed by culture or fermentation conditions. Unfortunately, the nonsporogenic yeast C. rugosa does not utilize the universal codon CTG for leucine; therefore, most of the CTG codons were converted to universal serine triplets by site‐directed mutagenesis to gain expression of functional lipase in heterologous hosts. Recombinant expressions by multiple‐site mutagenesis or complete synthesis of the lipase gene are other possible ways of obtaining pure and different CRL isoforms, in addition to culture engineering. Protein engineering of purified CRL isoforms allows the tailoring of enzyme function. This involves computer modeling based on available 3‐D structures of lipase isoforms. Lid swapping and DNA shuffling techniques can be used to improve the enantioselectivity, thermostability, and substrate specificity of CRL isoforms and increase their biotechnological applications. Lid swapping can result in chimera proteins with new functions. The sequence of the lid can affect the activity and specificity of recombinant CRL isoforms. Candida rugosa lipase is toxicologically safe for food applications. Protein engineering through lid swapping and rationally designed site‐directed mutagenesis will continue to lead to the production of CRL isoforms with improved catalytic power, thermostability, enantioselectivity, and substrate specificity, while providing evidence for the mechanisms of actions of the various isoforms.
Effects of specific conjugated linoleic acid isomers on growth characteristics in obese Zucker rats
Authors: Sanders Sara R. , Teachey Mary K. , Ptock Arne , Kraemer Klaus , Hasselwander Oliver , Henriksen Erik J. , Baumgard Lance H.
Published: November 2006

Growing female obese Zucker (fa/fa) rats were treated (via intragastric gavage) for 21 d with either a (i) vehicle [corn oil; 0.9 g/kg body weight (BW)], (ii) CLA mixture [50∶50; trans‐10,cis‐12 and cis‐9,trans‐11 CLA], (iii) cis‐9,trans‐11 CLA, or (iv) trans‐10,cis‐12 CLA (CLA treatments at 1.5 g CLA/kg BW). Compared with controls, average daily gain (g/d) was reduced 24 and 44% by the CLA mixture and trans‐10,cis‐12 CLA, respectively There was no treatment effect on average whole‐body (minus heart and liver) composition (dry matter basis): fat (70.2%), protein (21.0%), and ash (4.3%). Compared with animals treated with cis‐9,trans‐11 CLA, obese Zucker rats treated with trans‐10,cis‐12 and the CLA mixture had 7.8% more carcass water. Treatment had no effect on heart or liver weights or on heart or liver weights as a percentage of body weight, but compared with the other treatments trans‐10,cis‐12 CLA increased liver lipid contentby 33%. Hepatic lipid ratios of 16∶1/16∶0 and 18∶1/18∶0 (a proxy for Δ9‐desaturase capability) were not affected by treatment (0.1 and 0.6, respectively). Simlar to previous reports, CLA increased hepatic lipid content and altered both liver and carcass FA composition (i.e., reduced arachidonic acid content), but the ability of CLA to manipulate body composition in obese Zucker rats remains questionable.
Phospholipase D and phosphatidate phosphohydrolase activities in rat cerebellum during aging
Authors: Pasquaré S. J. , Salvador G. A. , Giusto N. M.
Published: November 2006

Aging is a process that affects different organs, of which the brain is particularly susceptible. PA and DAG are central intermediates in the phosphoglyceride as well as in the neutral lipid biosynthetic pathway, and they have also been implicated in signal transduction. Phospholipase D (PLD) and phosphatidate phosphohydrolase (PAP) are the enzymes that generate PA and DAG. The latter can be transformed into MAG by diacylglycerol lipase (DGL). In the present study, we examine how aging modulates the PLD, PAP, and DGL isoforms in cerebellar subcellular fractions from 4‐(adult),28‐, and 33‐mon‐old (aged) rats. Pl‐4,5‐bisphosphonate (PIP2)‐dependent PLD, PAP1, and DGL1 were distributed in different percentages in all cerebellum subcellular fractions. On the other hand, PAP2 and DGL2 activities were observed in all subcellular fractions except in the cytosolic fraction. Aging modified the enzyme distribution pattern. In addition, aging decreased nuclear (45%), mitochondrial‐synaptosomal (55%), and cytosolic (71%) PAP1 activity and increased (28%) microsomal PAP1 activity. DGL1 activity was decreased in nuclear (85%) and mitochondrial‐synaptosomal (63%) fractions by aging. On the other hand, PIP2‐dependent PLD activities were increased in the mitochondrial‐synaptosomal fraction. PAP2 and DGL2 were increased in the microsomal fraction by 87 and 114%, respectively, and they were decreased in the nuclear fraction. The changes observed in cerebellum PAP1 and DGL1 activities from aged rats with respect to adult rats could be related to modifications in lipid metabolism. Differential PA metabolization during aging through PIP2‐dependent PLD/PAP2/DGL2 activities could be related to alterations in the neural signal transduction mechanisms.
Isolation and structures of two divinyl ether fatty acids from Clematis vitalba
Authors: Hamberg Mats
Published: November 2006

[1‐14C]Linolenic acid was incubated with a homogenate of leaves of Clematis vitalba, a plant belonging to the Ranunculaceae family. Analysis of the reaction product by reversed‐phase high‐performance liquid radiochromatography demonstrated the presence of the following labeled oxylipins: 12‐oxo‐10,15(Z)‐phytodienoic acid, 9(S‐hydroxy‐10(E),12(Z),15(Z)‐octadecatrienoic acid, ω5(Z)‐etherolenic acid, and 9‐[1′(E),3′(Z),6′(Z)‐nonatrienyloxy]‐8(Z)‐nonenoic acid [8(Z)‐colnelenic acid]. The last compound was a new divinyl ether FA, and an analogous compound, i.e., 9‐[1′(E),3′(Z)‐nonadienyloxy]‐8(Z)‐nonenoic acid [8(Z‐colneleic acid], was obtained following incubation of linoleic acid with the Clematis homogenate. Structures of the two divinyl ethers were assigned by spectral and chromatographic comparison with authentic compounds prepared synthetically using previously described methodology. Separate incubation of the 9‐ and 13‐hydroperoxides of linolenic acid demonstrated that the first hydroperoxide served as the precursor of 8(Z)‐colnelenic acid and indicated the presence in C. vitalba of a new divinyl ether synthase acting on 9‐lipoxygenase‐generated hydroperoxides. A close structural relationship between this enzyme and the well‐studied divinyl ether synthase in the potato and tomato seems likely.
Geranium sanguineum (Geraniaceae) seed oil: A new source of petroselinic and vernolic acid
Authors: Tsevegsuren N. , Aitzetmuller K. , Vosmann K.
Published: November 2006

The occurrent of petroselinic acid (18∶1Δ6cis) in seed oils was believed to be limited to the Umbelliferae or Apiaceae, and a few other members of the Umbelliflorae. A major occurrence of petroselinic acid outside the Umbelliflorae must therefore be regarded as highly unusual and surprising. The seed oil of Geranium sanguineum, a member of the family Geraniaceae, has now been found to contain petroselinic and vernolic acids as major FA in its seed oil TAG. These unusual FA have not been reported previously as constituents of Geraniaceae seed oils. The structure and composition of the seed oil FA from G. sanguineum were determined by combined use of chromatographic (TLC, capillary GLC) and spectroscopic (IR, GC‐MS) techniques. The double‐bond position in petroselinic acid was located unambiguously by the characteristic mass fragmentation of its dimethyldisulfide (DMDS) adduct. The epoxy FA was identified as vernolic acid by co‐chromatography and by the mass fragments formed during GC‐MS of the products of the epoxy ring‐opening reaction with BF3 in methanol.
Reaction of mono‐epoxidized conjugated linoleic acid ester with boron trifluoride etherate complex
Authors: Jie Marcel S. F. Lie Ken , Lam Corey N. W.
Published: November 2006

The reaction of methyl 11, 12‐E‐epoxy‐9Z‐octadecenoate (1) with boron trifluoride etherate furnished a mixture of methyl 12‐oxo‐10E‐octadecenoate (3a) and methyl 11‐oxo‐9E‐octadecenoate (3b) in 66% yield. Methyl 9, 10‐Z‐epoxy‐11 E‐octadecenoate (2) with boron trifluoride etherate furnished a mixture of methyl 9‐oxo‐10 E‐octadecenoate (4a, 45%) and methyl 10‐oxo‐11 E‐octadecenoate (4b, 19%). A plausible mechanism is proposed for these reactions, which involves the attack on the epoxy ring system by BF3, followed by deprotonation, oxo formation, and double bond migration to give a mixture of two positional α,β‐unsaturated C18 enone ester derivatives (3a/3b, 4a/4b). The structures of these C18 enone ester derivatives (3a/3b, 4a/4b) were identified by a combination of NMR spectroscopic and mass spectrometric analyses.
Successful utilization of lyophilized lipoprotein(a) as a biological reagent
Authors: Scanu Angelo M. , Hinman Janet , Pfaffinger Ditta , Edelstein Celina
Published: November 2006

Lipoprotein(a) [Lp(a)] represents a class of lipoprotein particles having as a protein moiety apoB‐100 linked by a single disulfide bond to apolipoprotein(a) [apo(a)], a multikringle structure with a high degree of homology with plasminogen. A recognized feature of Lp(a) is its instability on storage caused by attendant protein and lipid modifications that affect the structural, functional, and immunological properties of this lipoprotein. Here we present data showing that, under appropriate conditions of cryopreservation, Lp(a) retains the properties of the freshly isolated product, and we provide examples supporting the stability of this cryopreserved product as a primary standard in immunoassay settings and in cell culture systems.
A short, concise route to diphosphatidylglycerol (Cardiolipin) and its variants
Authors: Krishna U. Murali , Ahmad Moghis U. , Ali Shoukath M. , Ahmad Imran
Published: November 2006

A new approach is described for the synthesis of the cardiolipin family of phospholipids that uses phosphonium salt methodology. The method involves the reaction of 2‐O‐protected glycerol with a trialkyl phosphite derived from 1,2‐diacyl‐sn‐glycerol in the presence of pyridinium bromide perbromide and triethylamine to afford the phosphoric triesters. The synthesis involves three steps and allows the preparation of a wide range of cardiolipins with different substitution patterns and chain lengths, including unsaturated derivatives. The use of inexpensive protecting groups and the ease of purification facilitate this synthetic route and allow its scale‐up in a higher overall yield (72%) than the literature methods.
The age‐related decline in intestinal lipid uptake is associated with a reduced abundance of fatty acid‐binding protein
Authors: Woudstra Trudy D. , Drozdowski Laurie A. , Wild Gary E. , Clandinin M. T. , Agellon Luis B. , Thomson Alan B. R.
Published: November 2006

Aging is associated with changes in the absorptive capacity of the small intestine. We tested the hypotheses that (i) aging is associated with a decline in lipid absorption, and that (ii) this decreased lipid absorption is due to a decline in the abundance of mRNA and/or the enterocyte cytosolic intestinal FA‐binding protein (I‐FABP), the liver FA‐binding protein (L‐FABP), and the ileal lipid‐binding protein (ILBP). In vitro uptake studies were performed on Fischer 344 rats at ages 1, 9, and 24 mon. Northern blotting (L‐FABP, ILBP) and immunohistochemistry (I‐FABP, ILBP) were performed. Aging was associated with decreased animal weights, but the surface area of the intestine was not significantly altered with age. The rates of ileal uptake of 16∶0, 18∶0, 18∶1, and 18∶2 were reduced by greater than 50% with aging when expressed on the basis of mucosal weight. This decline was not associated with reduced expression of mRNA for L‐FABP or ILBP but was associated with a 50% decrease in the abundance of I‐FABP and a 40% decrease in the abundance of ILBP. Thus, the decrease with aging in the ileal uptake of some FA when rates were expressed on the basis of mucosal weight was associated with a reduced abundance of I‐FABP and ILBP.
Conjugated linoleic acid isomer effects in atherosclerosis: Growth and regression of lesions
Authors: Kritchevsky David , Tepper Shirley A. , Wright Scott , Czarnecki Susanne K. , Wilson Thomas A. , Nicolosi Robert J.
Published: November 2006

Conjugated linoleic acid (CLA), a mixture of positional and geometric isomers of octadecadienoic acid, has been shown to inhibit experimentally induced atherosclerosis in rabbits and also to cause significant regression of pre‐established atheromatous lesions in rabbits. The two major CLA isomers (cis9,trans11 and trans10,cis12), now available at 90% purity, have been tested individually for their anti‐atherogenic or lesion regression potency. The two major isomers and the mixture were fed for 90 d to rabbits fed 0.2% cholesterol. Atherosclerosis was inhibited significantly by all three preparations. The two CLA isomers and the isomer mix were also fed (1.0%) as part of a cholesterol‐free diet for 90 d to rabbits bearing atheromatous lesions produced by feeding an atherogenic diet. A fourth group was maintained on a cholesterol‐free diet. On the CLA‐free diet atherosclerosis was exacerbated by 35%. Reduction of severity of atheromatous lesions was observed to the same extent in all three CLA‐fed groups. The average reduction of severity in the three CLA‐fed groups was 26±2% compared with the first control (atherogenic diet) and 46±1% compared with the regression diet. Insofar as individual effects on atherosclerosis were concerned, there was no difference between the CLA mix and the cis9,trans11 and trans10,cis12 isomers. They inhibit atherogenesis by 50% when fed as a component of a semipurified diet containing 0.2% cholesterol; and when fed as part of a cholesterol‐free diet, they reduce established lesions by 26%. Reduction of atheromata to the observed extent by dietar means alone is noteworthy.
Gestational age and birth weight in relation to n−3 fatty acids among inuit (Canada)
Authors: Lucas Michel , Dewailly Éric , Muckle Gina , Ayotte Pierre , Bruneau Suzanne , Gingras Suzanne , Rhainds Marc , Holub Bruce J.
Published: November 2006

Seafood consumption during pregnancy carries both benefits (high n−3 FA intake) and risks (exposure to environmental contaminants) for the developing fetus. We determined the impacts of marine n−3 FA and environmental contaminants on gestational age (GA) of Nunavik women and the anthropometric characteristics of their newborns. FA and contaminant (polychlorinated biphenyls and mercury) concentrations were measured in cord plasma of Nuvavik newborns (n=454) and compared with those of a group of newborns (n=29) from southern Québec. Data were collected from hospital records and birth certificates. In Nunavik newborns, arachidonic acid (AA) was two times lower (P<0.0001), whereas DHA concentration, the Σn−3/Σn−6 ratio, and the percentage of n−3 highly unsaturated FA (HUFA) (of the total HUFA) were three times higher (P<0.0001) compared with southern Québec newborns. After controlling for confounders, GA and birth weight were higher by 5.4 d [95% confidence interval (CI): 0.7–10.1] and 77 g (95% CI: −64 to 217) in the third tertile of percentage of n−3 HUFA (of the total HUFA) as compared with the first tertile. There was no evidence that contaminants had negative effects on GA or birth weight. In this seafood‐eating population, an increase in the proportion of n−3 HUFA (of the total HUFA), measured in umbilical cord plasma phospholipids, was associated with a significantly longer GA.
Supplementation and delivery of n−3 fatty acids through spray‐dried milk reduce serum and liver lipids in rats
Authors: Ramaprasad T. R. , Baskaran V. , Sambaiah K. , Lokesh B. R.
Published: November 2006

Indian diets comprising staples such as cereals, millets, and pulses provide 4.8 energy % from linoleic acid (18∶2n−6) but fail to deliver adequate amounts of n−3 FA. Consumption of long‐chain n−3 PUFA such as EPA (20∶5n−3) and DHA (22∶6n−3) is restricted to those who consume fish. The majority of the Indian population, however, are vegetarians needing additional dietary sources of n−3 PUFA. The present work was designed to use n−3 FA‐enriched spray‐dired milk powder to provide n−3 FA. Whole milk was supplemented with linseed oil to provide α‐linolenic acid (LNA, 18∶3n−3), with fish oil to provide EPA and DHA, or with groundnut oil (GNO), which is devoid of n−3 PUFA, and then spray‐dired. Male Wistar rats were fed the spray‐dired milk formulations for 60 d. The rats given formulations containing n−3 FA showed significant increases (P<0.001) in the levels of LNA or EPA/DHA in the serum and in tissue as compared with those fed the GNO control formulation. Rats fed formulations containing n−3 FA had 30–35% lower levels of serum total cholesterol and 25–30% lower levels of serum TAG than control animals. Total cholesterol and TAG in the livers of rats fed the formulations containing n−3 FA were lower by 18–30% and 11–18%, respectively, compared with control animals. This study showed that spray‐dried milk formulations supplemented with n−3 FA are an effective means of improving dietary n−3 FA intake, which may decrease the risk factors associated with cardiovascular disease.
Diet and lipoprotein oxidation: Analysis of oxidized triacylglycerols in pig lipoproteins
Authors: Suomela Jukka‐Pekka , Ahotupa Markku , Sjövall Olli , Kurvinen Juha‐Pekka , Kallio Heikki
Published: November 2006

Oxidized lipoproteins have a recognized role in atherogenesis, but molecular‐level research on oxidized lipids in lipoproteins and the effect of diet on these molecules have been limited. In the present study, the effects of three sunflower seed oil diets differing in oxidation levels (PV in oils 1, 84, and 223 mequiv O2/kg) on lipoprotein lipid oxidation in growing pigs were investigated. The emphasis was on the investigation of oxidized TAG molecules found in chylomicrons and VLDL. A method based on RP‐HPLC and electrospray ionization‐MS was used for the analysis of oxidized TAG molecules. The baseline diene conjugation method was used for the estimation of in vivo levels of lipoprotein lipid oxidation. Several oxidized TAG structures were found in the samples. These products consisted of TAG molecules with a hydroxy, an epoxy, or a keto group attached to a FA, and of TAG molecules containing an aldehyde structure derived from a FA. The lipoprotein lipids and TAG were more oxidized in the pigs fed on the most oxidized oil compared with those fed on nonoxidized oil. Oxidation of dietary fat was reflected in the lipoprotein oxidation. New, detailed information on oxidized TAG molecules of chylomicrons and VLDL was obtained.
Synthesis, spectroscopic, and biological studies of novel estolides derived from anticancer active 4‐O‐podophyllotoxinyl 12‐hydroxyl‐octadec‐Z‐9‐enoate
Authors: Mustafa Jamal , Khan Shabana I. , Ma Guoyi , Walker Larry A. , Khan Ikhlas A.
Published: November 2006

Podophyllotoxin is a well‐known natural antitumor agent with severe side effects, which led us to synthesize its numerous analogs in search of product(s) of improved therapeutic potential. Here, we report an efficient method for the synthesis of a series of 4‐O‐podophyllotoxin estolides with spectral characteristics and their biological studies. The OH of a known molecule, 4‐O‐podophyllotoxinyl 12‐hydroxyl‐octadec‐Z‐9‐enoate 2, was coupled with the carboxylic groups of different FA with the help of dicyclohexylcarbodiimide and dimethyl aminopyridine (catalyst) to produce high yields of their respective C4α‐estolides 3–11. Spectroscopic techniques, particularly 1H and 13CNMR, proved to be suitable tools to characterize the new compounds. These molecules of greater lipophilic character were tested for their in vitro cytotoxicity against four human solid tumors, one human leukemia cell, and one noncancerous cell. Compounds 4–6 and 11 showed moderate antileukemic activity; unexpectedly, none were found to be active against solid tumors. Estolides were also investigated for their in vitro activity against tubulin and topoisomerase II proteins. All the compounds showed inhibition of the catalytic activity of topoisomerase II, whereas 6–8 also inhibited tubulin polymerization. These results suggest the need for further screening of these molecules against a larger panel of cancerous cells.
Antibacterial and xanthine oxidase inhibitory cerebrosides from Fusarium sp. IFB‐121, and endophytic fungus in Quercus variabilis
Authors: Shu R. G. , Wang F. W. , Yang Y. M. , Liu Y. X. , Tan R. X.
Published: November 2006

Two antibacterial and xanthine oxidase inhibitory cerebrosides, one of which is chemically new, were characterized from the chloroform‐methanol (1∶1) extract of Fusarium sp. IFB‐121, an endophytic fungus in Quercus variabilis. By means of chemical and spectral methods [IR, electrospray ionization MS (ESI‐MS), tandem ESI‐MS, 1H and 13C NMR, distortionless enhancement by polarization transfer, COSY, heteronuclear multiple‐quantum coherence, heteronuclear multiple‐bond correlation, and 2‐D nuclear Overhauser effect correlation spectroscopy], the structure of the new metabolite named fusaruside was established as (2S,2′R,3R,3′E,4E,8E,10E)‐1‐O‐β‐d‐glucopyranosyl‐2‐N‐(2′‐hydroxy‐3′‐octadecenoyl)‐3‐hydroxy‐9‐methyl‐4,8,10‐sphingatrienine, and the structure of the other was identified as (2S,2′R,3R,3′E,4E,8E)‐1‐O‐β‐d‐glucopyranosyl‐2‐N‐(2′‐hydroxy‐3′‐octadecenoyl)‐3‐hydroxy‐9‐methyl‐4,8‐sphingadienine. Both new and known cerebrosides, although inactive to Trichophyton rubrum and Candida albicans, showed strong antibacterial activities against Bacillus subtilis, Escherichia coli, and Pseudomonas fluorescens, with their minimum inhibitory concentrations being 3.9, 3.9, and 1.9 μg/mL, and 7.8, 3.9, and 7.8 μg/mL, respectively. Furthermore, both metabolites were inhibitory to xanthine oxidase, with the IC50 value of fusaruside being 43.8±3.6 μM and the known cerebroside being 55.5±1.8 μM.
Quantitative determination of low density lipoprotein oxidation by FTIR and chemometric analysis
Authors: Lam Henry S. , Proctor Andrew , Nvalala John , Morris Manford D. , Smith W. Grady
Published: November 2006

This study was conducted to develop a quantitative FTIR spectroscopy method to measure LDL lipid oxidation products and determine the effect of oxidation on LDL lipid and protein. In vitro LDL oxidation at 37°C for 1 h produced a range of conjugated diene (CD) (0.14–0.26 mM/mg protein) and carbonyl contents (0.9–3.8 μg/g protein) that were used to produce calibration sets. Spectra were collected from the calibration set and partial least squares regression was used to develop calibration models from spectral regions 4000‐650, 3750‐3000, 1720‐1500, and 1180‐935 cm−1 to predict CD and carbonyl contents. The optimal models were selected based on their standard error of prediction (SEP), and the selected models were performance‐tested with an additional set of LDL spectra. The best models for CD prediction were derived from spectral regions 4000‐650 and 1180‐935 cm−1 with the lowest SEP of 0.013 and 0.013 mM/mg protein, respectively. The peaks at 1745 (cholesterol and TAG ester C=O stretch), 1710 (carbonyl C‐O stretch), and 1621 cm−1 (peptide C=O stretch) positively correlated with LDL oxidation. FTIR and chemometrics revealed protein conformation changes during LDL oxidation and provided a simple technique that has potential for rapidly observing structural changes in human LDL during oxidation and for measuring primary and secondary oxidation products.
Highly unsaturated fatty acid synthesis in vertebrates: New insights with the cloning and characterization of a Δ6 desaturase of Atlantic salmon
Authors: Zheng Xiaozhong , Tocher Douglas R. , Dickson Cathryn A. , Bell J. Gordon , Teale Alan J.
Published: November 2006

Fish are an important source of the n−3 highly unsaturated fatty acids (HUFA), eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids that are crucial to the health of higher vertebrates. The synthesis of HUFA involves enzyme‐mediated desaturation, and a Δ5 fatty acyl desaturase cDNA has been cloned from Atlantic salmon (Salmo salar) and functionally characterization of a Δ6 fatty acyl desaturase of Atlantic salmon and describe its genomic structure, tissue expression, and nutritional regulation. A salmon genomic library was screened with a salmon Δ5 desaturase cDNA and positive recombinant phage isolated and subcloned. The full‐length cDNA for the putative fatty acyl desaturase was shown to comprise 2106 bp containing an open reading frame of 1365 bp specifying a protein of 454 amino acids (GenBank accession no. AY458652). The protein sequence included three histidine boxes, two transmembrane regions, and an N‐terminal cytochrome b5 domain containing the heme‐binding motif HPGG, all of which are characteristic of microsomal fatty acid desaturases. Functional expression showed that this gene possessed predominantly Δ6 desaturase activity. Screening and sequence analysis of the genomic DNA of a single fish revealed that the Δ6 desaturase gene constituted 13 exons in 7965 bp of genomic DNA. Quantitative real‐time PCR assay of gene expression in Atlantic salmon showed that both Δ6 and Δ5 fatty acyl desaturase genes, and a fatty acyl elongase gene, were highly expressed in intestine, liver, and brain, and less so in kidney, heart, gill, adipose tissue, muscle, and spleen. Furthermore, expression of both Δ6 and Δ5 fatty acyl desaturase genes in intestine, liver, red muscle, and adipose tissue was higher in salmon fed a diet containing vegetable oil than in fish fed a diet containing fish oil.
β‐oxidation capacity of red and white muscle and liver in Atlantic salmon (Salmo salar L.)—Effects of increasing dietary rapeseed oil and olive oil to replace capelin oil
Authors: Stubhaug Ingunn , Frøyland Livar , Torstensen Bente E.
Published: November 2006

Post‐smolt Atlantic salmon (Salmo salar) were fed six diets in which capelin oil was replaced with 0, 25, 50, 75, or 100% rapeseed oil (RO; low‐erucic acid) or 50% olive oil (OO). The experimental diets were fed to single groups of Atlantic salmon for 42 wk, whereas the 100% capelin oil (0% RO) diet was fed in duplicate. The β‐oxidation capacity of palmitoyl‐CoA was determined, using a method optimized for salmon tissues, at the start of the experiment, after 21 wk (October), and after 42 wk (March) in red and white muscle and in liver. Red muscle showed the highest specific β‐oxidation capacity, but when expressed as total β‐oxidation capacity for the whole tissue, white muscle was the most important tissue for the β‐oxidation of FA. From the initial to the final sampling, the β‐oxidation capacity of white muscle increased significantly, whereas the β‐oxidation capacity in liver decreased significantly. After 22 wk, white muscle exhibited an increased β‐oxidation capacity when the dietary RO content was raised from 25 to 75%, with similar effects in red muscle and liver after 42 wk of feeding. The present results also show that the β‐oxidation capacity increased with an increase in fish size.
Restoration of depressed prostanoid‐induced ileal contraction in spontaneously hypertensive rats by dietary fish oil
Authors: Patten Glen S. , Adams Michael J. , Dallimore Julie A. , Rogers Paul F. , Topping David L. , Abeywardena Mahinda Y.
Published: November 2006

We have reported that dietary fish oil (FO) rich in n−3 PUFA modulates gut contractility. It was further demonstrated that the gut of spontaneously hypertensive rats (SHR) has a depressed contractility response to prostaglandins (PG) compared with normotensive Wistar‐Kyoto (WKY) rats. We investigated whether feeding diets supplemented with n−3 PUFA increased gut contractility and restored the depressed prostanoid response in SHR gut. Thirteen‐week‐old SHR were fed diets containing fat at 5 g/100 g as coconut oil (CO), lard, canola oil containing 10% (w/w) n−3 FA as α‐linolenic acid (18∶3n−3), or FO (as HiDHA®, 22∶6n−3) for 12 wk. A control WKY group was fed 5 g/100 g CO in the diet. As confirmed, the SHR CO group had a significantly lower gut response to PGE2and PGF2αcompared with the WKY CO group. Feeding FO increased the maximal contraction response to acetylcholine in the ileum compared with all diets and in the colon compared with lard, and restored the depressed response to PGE2and PGF2αin the ileum but not the colon of SHR. FO feeding also led to a significant increase in gut total phospholipid n−3 PUFA as DHA (22∶6n−3) with lower n−6 PUFA as arachidonic acid (20∶4n−6). Canola feeding led to a small increase in ileal EPA (20∶5n−3) and DHA and in colonic DHA without affecting contractility. However, there was no change in ileal membrane muscarinic binding properties due to FO feeding. This report confirms that dietary FO increases muscarinic‐ and eicosanoid receptor‐induced contractility in ileum and that the depressed prostanoid response in SHR ileum, but not colon, is restored by tissue incorporation of DHA as the active nutrient.