Comparative studies on individual isomeric 18:1 acids in cow, goat, and ewe milk fats by low-temperature high-resolution capillary gas-liquid chromatography.

The trans- as well as the cis-18:1 isomer profiles were established in cow, goat, and ewe cheese fats, with the assumption that these are representative of the corresponding milks. Argentation thin-layer chromatography was combined with low-temperature high-resolution gas-liquid chromatography on 100-m highly polar capillary columns, thus adding precision to earlier data for these species. Despite differences in the absolute content of trans-18:1 isomers between species, the relative profiles were essentially similar. Except for the minor trans delta6-delta8 group, all trans-18:1 isomers with their ethylenic bonds between positions delta4 and delta16 (including the resolved critical pair delta13/delta14) were separated and quantitated individually. As expected, vaccenic (trans delta9-18:1) acid was the main isomer, accounting for as much as 37 to 50% of the total fraction. It was observed that the goat trans-18:1 isomer profile was usually rather close to that of cows in winter (barn feeding), whereas that of the ewe shows a seasonal dependence. The trans-18:1 profile of ewe milk fats from this study resembles that of cows in the transition period between winter and summer (pasture) feeding. Regarding the cis-18:1 acid fraction, two isomers (oleic and cis-vaccenic acids) accounted for ca. 97% of that fraction for the three species, with the cis-delta12 isomer ranked third. The analytical procedure employed here appears a convenient alternative to oxidative-based procedures (generally ozonolysis), taking less time and alleviating some drawbacks of the latter procedure.

Effects of delta5 polyunsaturated fatty acids of maritime pine (Pinus pinaster) seed oil on the fatty acid profile of the developing brain of rats.

Conifer (pine) seeds are a potential source of dietary oils, but their safety and nutritional properties are not well established. Conifer seed oils differ from common edible vegetable oils in having a series of unusual polyunsaturated fatty acids (PUFA) with a polymethylene-interrupted (PMI) double bond system and a double bond at the delta5 position. A rat study was conducted to assess whether delta5 PMI-PUFA of conifer seeds could alter the levels of n-6 and n-3 long-chain polyunsaturated fatty acids (LC-PUFA) in mothers' milk and the developing brain of fetuses and pups. Feeding maritime pine (Pinus pinaster) seed oil (MPO) diet with a delta5 PMI-PUFA content of 1.4 g/100 g throughout pregnancy and lactation resulted in a large incorporation of delta5 PMI-PUFA in mothers' milk (5.1 +/- 0.5% of total fatty acids). The fetus (17 d old) and pup (22 d) brains, however, accumulated very little (0.6 and 0.4% of total fatty acids, respectively) delta5 PMI-PUFA. Mother's milk and pup's brain of the MPO group contained normal levels of 20:4n-6, 22:4n-6, and 20:5n-3 compared to a reference group of rats fed a fat blend of sunflower, high-oleic sunflower, and canola oils. The level of 22:6n-3, however, was slightly but significantly (P < 0.05) higher in milk and pup brain of the MPO group. These results show that delta5 PMI-PUFA of MPO exert no negative effect on the levels of n-6 and n-3 LC-PUFA in rat brain during its early development.

Fatty acid composition of Pinaceae as taxonomic markers.

Following our previous review on Pinus spp. seed fatty acid (FA) compositions, we recapitulate here the seed FA compositions of Larix (larch), Picea (spruce), and Pseudotsuga (Douglas fir) spp. Numerous seed FA compositions not described earlier are included. Approximately 40% of all Picea taxa and one-third of Larix taxa have been analyzed so far for their seed FA compositions. Qualitatively, the seed FA compositions in the three genera studied here are the same as in Pinus spp., including in particular the same delta5-olefinic acids. However, they display a considerably lower variability in Larix and Picea spp. than in Pinus spp. An assessment of geographical variations in the seed FA composition of P. abies was made, and intraspecific dissimilarities in this species were found to be of considerably smaller amplitude than interspecific dissimilarities among other Picea species. This observation supports the use of seed FA compositions as chemotaxonomic markers, as they practically do not depend on edaphic or climatic conditions. This also shows that Picea spp. are coherently united as a group by their seed FA compositions. This also holds for Larix spp. Despite a close resemblance between Picea and Larix spp. seed FA compositions, principal component analysis indicates that the minor differences in seed FA compositions between the two genera are sufficient to allow a clear-cut individualization of the two genera. In both cases, the main FA is linoleic acid (slightly less than one-half of total FA), followed by pinolenic (5,9,12-18:3) and oleic acids. A maximum of 34% of total delta5-olefinic acids is reached in L. sibirica seeds, which appears to be the highest value found in Pinaceae seed FA. This apparent limit is discussed in terms of regio- and stereospecific distribution of delta5-olefinic acids in seed triacylglycerols. Regarding the single species of Pseudotsuga analyzed so far (P. menziesii), its seed FA composition is quite distinct from that of the other two genera, and in particular, it contains 1.2% of 14-methylhexadecanoic (anteiso-17:0) acid. In the three genera studied here, as well as in most Pinus spp., the C18 delta5-olefinic acids (5,9-18:2 and 5,9,12-18:3 acids) are present in considerably higher amounts than the C20 delta5-olefinic acids (5,11-20:2 and 5,11,14-20:3 acids).

A new delta 7-polyunsaturated fatty acid in Taxus spp. Seed lipids, dihomotaxoleic (7,11-20:2) acid.

A previously undescribed fatty acid, all-cis 7,11-20:2 (dihomotaxoleic acid, DHT), has been characterized by gas chromatography-mass spectrometry as being present (approximately 0.1%) in seed oils of two Taxaceae containing high levels (11-16%) of taxoleic acid (all-cis 5,9-18:2). This compound was absent from oils of 10 other conifer genera, as well as from one member of Taxaceae containing very low amounts of taxoleic acid, suggesting that DHT is a taxoleic acid elongation product.

Regiospecific analysis of conifer seed triacylglycerols by gas-liquid chromatography with particular emphasis on delta5-olefinic acids.

Dibutyroyl derivatives of monoacylglycerols (DBMAG) from conifer seed oil triacylglycerols (TAG) were prepared by partial deacylation of TAG with ethylmagnesium bromide followed by diesterification with n-butyryl chloride. The resulting mixtures were analyzed by gas-liquid chromatography (GLC) with a 65% phenylmethyl silicon open tubular fused-silica capillary column operated under optimal conditions and separated according to both their fatty acid structures and their regiospecific distribution. Seed oils of 18 species from 5 conifer families (Pinaceae, Taxaceae, Cupressaceae, Cephalotaxaceae, and Podocarpaceae) were analyzed. The chromatograms showed a satisfactory resolution of DBMAG containing palmitic (16:0), stearic (18:0), taxoleic (cis-5,cis-9 18:2), oleic (cis-9 18:1), cis-vaccenic (cis-11 18:1), pinolenic (cis-5,cis-9,cis-12 18:3), linoleic (cis-9,cis-12 18:2), alpha-linolenic (cis-9,cis-12,cis-15 18:3), and an almost baseline resolution of DBMAG containing gondoic (cis-11 20:1), cis-5,cis-11 20:2, sciadonic (cis-5,cis-11,cis-14 20:3), dihomolinoleic (cis-11,cis-14 20:2), juniperonic (cis-5,cis-11,cis-14,cis-17 20:4), and dihomo-alphalinolenic (cis-11,cis-14,cis-17 20:3) acids. We have observed that results for Pinus pinaster and P. koraiensis seed oils obtained with this new simple method compared favorably with literature data established with other usual regiospecific analytical techniques. Delta5-olefinic acids are esterified mainly at the external positions of the glycerol backbone in all cases, in agreement with data obtained by other methodologies allowing validation of the GLC regiospecific method. To date, 45 gymnosperm species (mostly Coniferophytes) from 21 genera belonging to 9 families have been analyzed, all of them showing a definite enrichment of delta5-olefinic acids in the external positions of TAG. These fatty acids (FA), with one exception only, represent between approximately 2 and 8% of FA esterified to the internal positions. For some species, i.e., P. koraiensis and P. pinaster, this asymmetrical distribution was established by at least three analytical procedures and confirmed by stereospecific analysis of their seed TAG.

Trans- and cis-octadecenoic acid isomers in the hump and milk lipids from Camelus dromedarius.

The distribution profiles of individual trans- as well as cis-18:1 isomers from the fat prepared from the hump adipose tissue and the milk from Camelus dromedarius (the single-humped Arabian species) are described. Gas-liquid chromatography on two capillary columns with different polarities and lengths were used for this purpose in combination with argentation thin-layer chromatography. A comparison of the profiles established is made with that of true ruminant fats. In the fats from the dromedarius as well as from true ruminants, the trans-18:1 isomers have their ethylenic bonds in all positions between delta4 and delta16. The prominent trans isomer is the 11-18:1 (vaccenic) acid in all species, and the complete distribution profiles are quite similar. Concerning the cis isomers, the prominent isomer is oleic acid, followed by cis-vaccenic acid, as in true ruminant fats. Other cis isomers encompass the delta6-8 and the delta12 to delta15 isomers. Camelidae (suborder Tylopoda) and Bovidae (suborder Ruminantia) have evolved independently since the Eocene, that is for approximately 50 million years. Despite this considerable period, and the profound differences in anatomy, morphology, physiology, ecological and dietary habits between the extant species of these suborders, the rumen microflora has continued to synthesize the same trans- and cis-octadecenoic acid isomers, in comparable proportions, at least as deduced from their composition profiles. We conclude that the trans-18:1 acid profile is not intrinsically species-dependent, but it can be affected by the nature and the proportions of dietary unsaturated fatty acids that themselves depend on the feed, and that may be species-specific.

Effects of dietary maritime pine seed oil on lipoprotein metabolism and atherosclerosis development in mice expressing human apolipoprotein B.

BACKGROUND: Conifer seeds are used for food preparation in several countries. Aim of the study To assess the lipid-lowering and antiatherogenic properties of maritime pine (Pinuspinaster) seed oil. METHODS: The effects of maritime pine oil supplementation (20% w/w) for 2 weeks were compared to those of coconut and sunflower oil in mice expressing human apolipoprotein B (hApoB). Atherosclerosis lesion development was measured in hApoB mice fed 1.25% (w/w) cholesterol and 0.05% (w/w) sodium cholate and either coconut, sunflower or maritime pine oil (20% w/w) for 8 weeks. RESULTS: After 2 weeks of dietary treatment, plasma cholesterol (p < 0.0001), triglyceride (p < 0.0003), phospholipid (p < 0.0001) and apolipoprotein B (p < 0.0001) levels were lower in mice supplemented with maritime pine oil than in those treated with coconut oil. These effects were accounted for by a lowering of LDL-cholesterol, LDL-phospholipids and LDL-triglycerides, as well as a decrease in HDL-cholesterol and HDL-phospholipids. After 8 weeks of dietary treatment cholesterol and cholate, the mean area of aortic lesions was not statistically different between fat groups. CONCLUSIONS: Feeding maritime pine oil is associated with major changes of lipid and lipoprotein levels in hApoB mice. However, in the long term, maritime pine oil has no preventive effect on cholesterol-induced aortic lesion development in hApoB mice.

Study of individual trans- and cis-16:1 isomers in cow, goat, and ewe cheese fats by gas-liquid chromatography with emphasis on the trans-delta3 isomer.

Low-temperature gas-liquid chromatography (GLC) was applied to study the distribution profiles of isomeric trans- and cis-hexadecenoic acids in ruminant (cow, goat, and ewe) milk fat after their fractionation by argentation thin-layer chromatography (Ag-TLC). The fat was extracted from cheeses (12 samples of each species), the most common foods made with goat and ewe milks. The predominant trans-16:1 isomer is palmitelaidic acid (the delta9 isomer), but it does not exceed one-third of the total group, which itself represents 0.17% (cow), 0.16% (goat), and 0.26% (ewe) of the total fatty acids. The trans-delta3 16:1 isomer, which is reported for the first time in ruminant lipids and which likely comes from the animals' feed, is present at a level of ca. 10% of the trans-16:1 acid group. Otherwise, all isomers with their ethylenic bond between positions delta4 and delta14 are observed in the three species studied, roughly showing the same relative distribution pattern. Quantitatively, the trans-16:1 isomers only represent ca. 5% of the sum of the trans-16:1 plus trans-18:1 isomers, and they appear of little importance in comparison. It is inferred from this and recent studies that some previously reported data that were established for consumption assessments dealt in fact mainly with iso-17:0 acid, which was confused with (and added to) trans-delta9 (palmitelaidic) acid; consequently, these results were large over-estimates. Regarding the cis-16:1 acids, the delta9 isomer is the prominent constituent as expected, but the second-most important isomer is the delta13 isomer. It does not appear that trans-16:1 isomers are from ruminant milk fats of great nutritional importance as compared with trans-18:1 isomeric acids. As for trans18:1 isomers, the combination Ag-TLC/GLC is a necessary procedure to quantitate trans-16:1 acids accurately and reliably. Ag-TLC allows removal of interfering branched 17:0 acids and cis-16:1 acids, and low-temperature GLC permits an accurate measurement of all individual isomers most of which with baseline resolution.

Effects of dietary maritime pine (Pinus pinaster)-seed oil on high-density lipoprotein levels and in vitro cholesterol efflux in mice expressing human apolipoprotein A-I.

Maritime pine (Pinus pinaster)-seed oil contains two Delta5 unsaturated polymethylene interrupted fatty acids (all cis-5,9, 12-18:3 and all cis-5,11,14-20:3 acids) one of which resembles eicosapentaenoic acid. The goal of the present study was to test whether maritime pine-seed oil consumption affects HDL and apolipoprotein (Apo) A-I levels as well as the ability of serum to promote efflux of cholesterol from cultured cells. To this end, wild type (WT) non-transgenic mice and transgenic mice expressing human ApoA-I (HuA-ITg) were fed on isoenergetic diet containing either 200 g maritime pine-seed oil/kg or 200 g lard/kg for 2 weeks. WT and HuA-ITg mice fed maritime pine-seed oil had lower cholesterol, HDL-cholesterol, LDL-cholesterol and HuA-ITg mice had lower human ApoA-I than those fed lard. The differences in cholesterol (P < 0.0001) and HDL-cholesterol (P < 0.003) levels between mice fed on the two diets were more pronounced in the HuA-ITg than in the WT mice. The ability of HuA-ITg serum to promote cholesterol efflux in cultured cells was greater (P < 0.008) than that of WT animals. However, the maritime pine-seed oil diet was associated with lower (P < 0.005) in vitro cholesterol efflux ability than the lard diet in both mice genotypes. This suggests a negative effect of the maritime pine-seed oil on reverse cholesterol transport. Cholesterol efflux was correlated with serum free or esterified cholesterol and phospholipid levels. The slope of the regression line was smaller in the HuA-ITg than in the WT mice indicating that overexpression of human ApoA-I reduces the negative impact of maritime pine-seed oil on cholesterol efflux. In conclusion, maritime pine-seed oil diet lowers HDL-cholesterol and diminishes in vitro cholesterol efflux. This potentially detrimental effect is attenuated by overexpression of human ApoA-I in mice.

Follow-up of the delta4 to delta16 trans-18:1 isomer profile and content in French processed foods containing partially hydrogenated vegetable oils during the period 1995-1999. Analytical and nutritional implications.

A survey of the total content of trans-18:1 acids and their detailed profile in French food lipids was conducted in 1995-1996, and 1999. For this purpose, 37 food items were chosen from their label indicating the presence of partially hydrogenated vegetable oils (PHVO) in their ingredients. The content as well as the detailed profile of these isomers was established by a combination of argentation thin-layer chromatography and gas liquid chromatography (GLC) on long polar capillary columns. With regard to the mean trans-18:1 acid contents of extracted PHVO, a significant decrease was observed between the two periods, i.e., from 26.9 to 11.8% of total fatty acids. However, only minor differences were noted in the mean relative distribution profiles of individual trans-18:1 isomers with ethylenic bonds between positions delta4 and delta16 for the two periods. The predominant isomer was delta9-18:1 (elaidic) acid, in the wide range 15.2-46.1% (mean, 27.9+/-7.2%) of total trans-18:1 acids, with the delta10 isomer ranked second, with a mean of 21.3% (range, 11.6 to 27.4%). The content of the unresolved delta6 to delta8 isomer group was higher than the delta11 isomer (vaccenic acid), representing on average 17.5 and 13.3%, respectively. Other isomers delta4, delta5, delta12, delta13/delta14, delta15, and delta16, were less than 10% each: 1.0, 1.6, 7.4, 7.1, 1.8, and 1.0%, respectively. However, considering individual food items, it was noted that none of the extracted PHVO were identical to one another, indicating a considerable diversity of such fats available to the food industry. A comparison of data for French foods with similar data recently established for Germany indicates that no gross differences occur in PHVO used by food industries in both countries. Estimates for the absolute mean consumption of individual isomers from ruminant fats and PHVO are made for the French population and compared to similarly reconstructed hypothetical profiles for Germany and North America. Differences occur in the total intake of trans-18:1 acids, but most important, in individual trans-18:1 isomer intake, with a particular increase of the delta6-delta8 to delta10 isomers with increasing consumption of PHVO. It is inferred from the present and earlier data that direct GLC of fatty acids is a faulty procedure that results (i) in variable underestimates of total trans-18:1 acids, (ii) in a loss of information as regards the assessment of individual isomeric trans-18:1 acids, and (iii) in the impossibility of comparing data obtained from human tissues if the relative contribution of dietary PHVO and ruminant fats is not known.

Comparison of maritime pine oil and fish oil effects on plasma lipoproteins in apolipoprotein E-deficient mice.

The goal of this study was to compare the lipid lowering properties of maritime pine and fish oils in apolipoprotein E-deficient (KOE) mice, an animal model of hyperlipidemia. KOE mice were supplemented with either lard, fish or maritime pine oil (10% w/w) for one month. Compared to lard, fish and maritime pine oils decreased cholesterol (-31% and -52% respectively) and phospholipid (-41 and -52%) levels and increased triglyceride (+182% and +123%) levels. These lipid changes resulted in an enrichment in triglycerides and a depletion in cholesterol of VLDL+IDL plasma fraction as compared to lard-fed mice. These findings suggest that VLDL-triglyceride lipolysis is impaired in KOE mice fed fish or maritime pine oil.

General characteristics of Pinus spp. seed fatty acid compositions, and importance of delta5-olefinic acids in the taxonomy and phylogeny of the genus.

The delta5-unsaturated polymethylene-interrupted fatty acid (delta5-UPIFA) contents and profiles of gymnosperm seeds are useful chemometric data for the taxonomy and phylogeny of that division, and these acids may also have some biomedical or nutritional applications. We recapitulate here all data available on pine (Pinus; the largest genus in the family Pinaceae) seed fatty acid (SFA) compositions, including 28 unpublished compositions. This overview encompasses 76 species, subspecies, and varieties, which is approximately one-half of all extant pines officially recognized at these taxon levels. Qualitatively, the SFA from all pine species analyzed so far are identical. The genus Pinus is coherently united--but this qualitative feature can be extended to the whole family Pinaceae--by the presence of delta5-UPIFA with C18 [taxoleic (5,9-18:2) and pinolenic (5,9,12-18:3) acids] and C20 chains [5,11-20:2, and sciadonic (5,11,14-20:3) acids]. Not a single pine species was found so far with any of these acids missing. Linoleic acid is almost always, except in a few cases, the prominent SFA, in the range 40-60% of total fatty acids. The second habitual SFA is oleic acid, from 12 to 30%. Exceptions, however, occur, particularly in the Cembroides subsection, where oleic acid reaches ca. 45%, a value higher than that of linoleic acid. Alpha-linolenic acid, on the other hand, is a minor constituent of pine SFA, almost always less than 1%, but that would reach 2.7% in one species (P. merkusii). The sum of saturated acids [16:0 (major) and 18:0 (minor) acids principally] is most often less than 10% of total SFA, and anteiso-17:0 acid is present in all species in amounts up to 0.3%. Regarding C18 delta5-UPIFA, taxoleic acid reaches a maximum of 4.5% of total SFA, whereas pinolenic acid varies from 0.1 to 25.3%. The very minor coniferonic (5,9,12,15-18:4) acid is less than 0.2% in all species. The C20 elongation product of pinolenic acid, bishomo-pinolenic (7,11,14-20:3) acid, is a frequent though minor SFA constituent (maximum, 0.7%). When considering C20 delta5-UPIFA, a difference is noted between the subgenera Strobus and Pinus. In the former subgenus, 5,11-20:2 and sciadonic acids are < or =0.3 and < or =1.9%, respectively, whereas in the latter subgenus, they are most often > or =0.3 and > or =2.0%, respectively. The highest values for 5,11-20:2 and sciadonic acids are 0.5% (many species) and 7.0% (P. pinaster). The 5,11,14,17-20:4 (juniperonic) acid is present occasionally in trace amounts. The highest level of total delta5-UPIFA is 30-31% (P. sylvestris), and the lowest level is 0.6% (P. monophylla). Uniting as well as discriminating features that may complement the knowledge about the taxonomy and phylogeny of pines are emphasized.

Arachidonic, eicosapentaenoic, and biosynthetically related fatty acids in the seed lipids from a primitive gymnosperm, Agathis robusta.

The fatty acid composition of the seeds from Agathis robusta, an Australian gymnosperm (Araucariaceae), was determined by a combination of chromatographic and spectrometric techniques. These enabled the identification of small amounts of arachidonic (5,8,11,14-20:4) and eicosapentaenoic (5,8,11,14,17-20:5) acids for the first time in the seed oil of a higher plant. They were apparently derived from gamma-linolenic (6,9,12-18:3) and stearidonic (6,9,12,15-18:4) acids, which were also present, via chain elongation and desaturation, together with other expected biosynthetic intermediates [bis-homo-gamma-linolenic (8,11,14-20:3) and bishomo-stearidonic (8,11,14,17-20:4) acids]. Also present were a number of C20 fatty acids, known to occur in most gymnosperm families, i.e., 5,11-20:2, 11,14-20:2 (bishomo-linoleic), 5,11,14-20:3 (sciadonic), 11,14,17-20:3 (bishomo-alpha-linolenic), and 5,11,14,17-20:4 (juniperonic) acids. In contrast to most other gymnosperm seed lipids analyzed so far, A. robusta seed lipids did not contain C18 delta5-desaturated acids [i.e., 5,9-18:2 (taxoleic), 5,9,12-18:3 (pinolenic), or 5,9,12,15-18:4 (coniferonic)]. These structures support the simultaneous existence of delta6- and delta5-desaturase activities in A. robusta seeds. The delta6-ethylenic bond is apparently introduced into C18 polyunsaturated acids, whereas the delta5-ethylenic bond is introduced into C20 polyunsaturated acids. A general metabolic pathway for the biosynthesis of unsaturated fatty acids in gymnosperm seeds is proposed. When compared to Bryophytes, Pteridophytes (known to contain arachidonic and eicosapentaenoic acids), and species from other gymnosperm families (without such acids), A. robusta appears as an "intermediate," with the C18 delta6-desaturase/C18-->C20 elongase/C20 delta5-desaturase system in common with the former subphyla, and the unsaturated C18-->C20 elongase/C20 delta5-desaturase system specific to gymnosperms. The following hypothetical evolutionary sequence for the C18 delta6/delta5-desaturase class in gymnosperm seeds is suggested: delta6 (initial)-->delta6/delta5 (intermediate)-->delta5 (final).

Delta5-olefinic acids in the seed lipids from four Ephedra species and their distribution between the alpha and beta positions of triacylglycerols. Characteristics common to coniferophytes and cycadophytes.

The fatty acid compositions of the seed lipids from four Ephedra species, E. nevadensis, E. viridis, E. przewalskii, and E. gerardiana (four gymnosperm species belonging to the Cycadophytes), have been established with an emphasis on delta5-unsaturated polymethylene-interrupted fatty acids (delta5-UPIFA). Mass spectrometry of the picolinyl ester derivatives allowed characterization of 5,9- and 5,11-18:2; 5,9,12-18:3; 5,9,12,15-18:4; 5,11-20:2; 5,11,14-20:3; and 5,11,14,17-20:4 acids. Delta5-UPIFA with a delta11-ethylenic bond (mostly C20 acids) were in higher proportions than delta5-UPIFA with a delta9 double bond (exclusively C18 acids) in all species. The total delta5-UPIFA content was 17-31% of the total fatty acids, with 5,11,14-20:3 and 5,11,14,17-20:4 acids being the principal delta5-UPIFA isomers. The relatively high level of cis-vaccenic (11-18:1) acid found in Ephedra spp. seeds, the presence of its delta5-desaturation product, 5,11-18:2 acid (proposed trivial name: ephedrenic acid), and of its elongation product, 13-20:1 acid, were previously shown to occur in a single other species, Ginkgo biloba, among the approximately 170 gymnosperm species analyzed so far. Consequently, Ephedraceae and Coniferophytes (including Ginkgoatae), which have evolved separately since the Devonian period (approximately 300 million yr ago), have kept in common the ability to synthesize C18 and C20 delta5-UPIFA. We postulate the existence of two delta5-desaturases in gymnosperm seeds, one possibly specific for unsaturated acids with a delta9-ethylenic bond, and the other possibly specific for unsaturated acids with a delta11-ethylenic bond. Alternatively, the delta5-desaturases might be specific for the chain length with C18 unsaturated acids on the one hand and C20 unsaturated acids on the other hand. The resulting hypothetical pathways for the biosynthesis of delta5-UPIFA in gymnosperm seeds are only distinguished by the position of 11-18:1 acid. Moreover, 13C nuclear magnetic resonance spectroscopy of the seed oil from two Ephedra species has shown that delta5-UPIFA are essentially excluded from the internal position of triacylglycerols, a characteristic common to all of the Coniferophytes analyzed so far (more than 30 species), with the possibility of an exclusive esterification at the sn-3 position. This structural feature would also date back to the Devonian period, but might have been lost in those rare angiosperm species containing delta5-UPIFA.

Pinus pinaster oil affects lipoprotein metabolism in apolipoprotein E-deficient mice.

The aim of the present study was to assess the antiatherogenic properties of Pinus pinaster (maritime pine) seed oil. To this end, the effects of P. pinaster oil supplementation on lipoprotein levels and atherosclerotic lesions were compared to those of lard or sunflower oil in apolipoprotein E-deficient mice. Plasma total cholesterol (P < 0.0001) and VLDL + intermediary density lipoprotein (IDL)-cholesterol (P < 0.0001) levels were lower in mice fed P. pinaster and sunflower oil than in those fed the lard diet. In contrast, triglycerides (P < 0.0001) and VLDL + IDL-triglycerides (P < 0.0001) levels were higher in mice fed P. pinaster oil than sunflower oil or lard. The VLDL + IDL lipid composition of apolipoprotein E-deficient mice fed P. pinaster oil was intermediate between that of lard-fed transgenic mice and that of wild-type mice fed nonpurified diet. Using the Triton WR1339 method, the fractional catabolic rate of plasma triglycerides was found to be lower in mice fed P. pinaster oil (P < 0.0001) than sunflower oil or lard diet, suggesting a defective clearance of triglycerides in the P. pinaster group. Finally, the susceptibility of triglyceride-rich lipoproteins to in vitro lipoprotein lipase-mediated lipolysis was lower in the P. pinaster oil-fed group than in the lard-fed group. Despite the differences in VLDL + IDL level and lipid composition, the surface areas of aortic atherosclerotic lesions were not significantly different among mice fed P. pinaster, sunflower or lard diets. In conclusion, the results of the present study indicated that feeding P. pinaster oil had no better preventive effect on aortic atherosclerotic lesion extension in apolipoprotein E-deficient mice than other saturated or polyunsaturated fats.

Effects of Pinus pinaster and Pinus koraiensis seed oil supplementation on lipoprotein metabolism in the rat.

The aim of the present study was to assess the effect of vegetal oils obtained from Pinus pinaster and P. koraiensis seeds on plasma lipoprotein levels and apolipoprotein (apo) gene expression in rats. These oils contain two particular fatty acids of the delta5-unsaturated polymethylene-interrupted fatty acid (delta5-UPIFA) family: all-cis-5,9,12-1 8:3 (pinolenic) and/or all-cis-5,11,14-20:3 (sciadonic) acids. Rats were fed for 28 d a diet containing 5% (w/w) oil supplement. Two control diets were prepared to match the fatty acid composition of P. pinaster or P. koraiensis oils with the exception of delta5-UPIFA, which were replaced by oleic acid. Pinus pinaster seed oil decreased serum triglycerides by 30% (P < 0.02), very low density lipoprotein (VLDL)-triglycerides by 40% (P < 0.01), and VLDL-cholesterol by 33% (P < 0.03). Pinus koraiensis seed oil decreased serum triglycerides by 16% [not statistically significant (ns)] and VLDL-triglycerides by 21% (ns). Gel permeation chromatography and nondenaturating polyacrylamide gel electrophoresis showed a tendency of high density lipoprotein to shift toward larger particles in pine seed oil-supplemented rats. Finally, P. pinaster seed oil treatment was associated with a small decrease of liver apoC-III (P < 0.02) but not in apoE, apoA-I, or apoA-II mRNA levels. The levels of circulating apo were not affected by pine seed oil supplementation. In conclusion, P. pinaster seed oil has a triglyceride-lowering effect in rats, an effect that is due to a reduction in circulating VLDL.

Essential fatty acids and their trans geometrical isomers in powdered and liquid infant formulas sold in Canada.

BACKGROUND: Animal and human studies have suggested that trans fatty acids might alter some physiological functions and adversely affect the growth and essential fatty acid balance of infants. In this context it is important to know the fatty acid composition, including the levels of trans isomers of oleic, linoleic and alpha-linolenic acids in infant formulas. METHODS: Ten liquid and fourteen powdered formulas for term infants were purchased from retail stores in Canada. The fatty acid composition of each formula was determined by capillary gas-liquid chromatography. RESULTS: All the formulas met the minimum content of 500 mg of linoleic acid/100 kcal formula (equivalent to 4.5% of energy) specified under current Canadian regulations. The formulas all met the minimum energy levels of 3% as linoleic acid and 0.7% as alpha-linolenic acid recommended recently by an ad hoc committee of Health Canada. However, in nine formulas, the proportion of linoleic acid was more than 20% of total fatty acids, and consequently, in five of them, the ratio of linoleic acid to alpha-linolenic acid exceeded the maximum ratio of 16:1 recommended by the ad hoc committee. Trans fatty acids were present in all the samples, and generally the liquid formulas displayed a higher total trans content (mean 1.9%, range 0.9-3.1%) than powdered formulas (mean 1.4%, range 0.6-2.5%). The amounts of trans isomers of linoleic and alpha-linolenic acids and the degree of isomerization of these fatty acids were also higher in liquid formulas than in powdered formulas. CONCLUSIONS: A few of the Canadian infant formulas would provide one-third of alpha-linolenic acid as trans geometric isomers.

The unusual occurrence of 14-methylhexadecanoic acid in Pinaceae seed oils among plants.

14-Methylhexadecanoic (14-MHD) acid has been identified in a sample of pine seed oil (Pinus contorta) by gas-liquid chromatography-mass spectrometry of its picolinyl ester derivative. Its identification (through its equivalent chain length) and its distribution in four conifer families have been checked. It occurred only in Pinaceae, where it was found in 72 species belonging to the genera Pinus, Abies, Cedrus, Tsuga, Pseudotsuga, Larix, and Picea, in the range 0.02-1.15%. 14-MHD acid could not be detected in the lipids of Taxaceae (Taxus baccata), Cupressaceae (Juniperus communis), or Taxodiaceae (Sciadopytis verticillata), even after a 10-fold concentration of the saturated acid fraction isolated by argentation thin-layer chromatography. It is concluded that Pinaceae, along with Ginkgo biloba seed lipids, are major exceptions in the plant kingdom with regard to 14-MHD acid, which otherwise occurs almost exclusively in lipids of animals and microorganisms. The biosynthesis and metabolic role of 14-MHD acid, which otherwise also occur in wood and leaf lipids, remain unknown.

Structural importance of the cis-5 ethylenic bond in the endogenous desaturation product of dietary elaidic acid, cis-5,trans-9 18:2 acid, for the acylation of rat mitochondria phosphatidylinositol.

When rats were fed elaidic (trans-9 18:1) acid at a high load in diets that were otherwise marginally or almost completely deficient in linoleic (cis-9,cis-12 18:2) acid, elaidic acid was desaturated to cis-5,trans-9 18:2 acid. This polymethylene-interrupted acid was then incorporated into most phospholipids from rat mitochondria, cardiolipin being an exception. Its level of esterification in phospholipids followed the increasing order: phosphatidylethanolamine < phosphatidylcholine < phosphatidylinositol (PI). The content of cis-5,trans-9 18:2 acid decreased in organs in the order liver > kidney > heart. The levels of cis-5,trans-9 18:2 acid increased in mitochondria phospholipids as the level of linoleic acid was lowered in the diet. In liver mitochondria PI, it reached 16% of total fatty acids. After hydrolysis of liver mitochondria PI with Naja naja phospholipase A2, we observed that elaidic acid was essentially esterified to position 1 at the expense of saturated acids, whereas cis-5,trans-9 18:2 acid was exclusively esterified to position 2, along with 20:3n-9 and 20:4n-6 acids. As a consequence, the sums of saturated and trans-9 18:1 acids on the one hand, and of 20:3n-9, 20:4n-6, and cis-5,trans-9 18:2 acids on the other hand, remained fairly constant in liver mitochondria PI (ca. 55 and 30%, respectively). Because trans-9 18:1 and cis-5,trans-9 18:2 acids differ only by the cis-5 ethylenic bond, which is also present in 20:3n-9 and 20:4n-6 acids, this distribution pattern indicates that the cis-5 double bond, rather than any other ethylenic bond, may be of major structural importance for channeling fatty acids to position 2 of PI.