Addition of triglycerides with arachidonic acid or docosahexaenoic acid to infant formula has tissue- and lipid class-specific effects on fatty acids and hepatic desaturase activities in formula-fed piglets

The effects of including triglycerides with arachidonic [20:4(n-6)] or docosahexaenoic acid [22:6(n-3)] in formula on plasma chylomicron, LDL and HDL, liver, heart, kidney and brain (n-6) and (n-3) fatty acids were investigated in formula-fed piglets. Piglets were fed formula with (in % total fatty acids) 20% 18:2(n-6) and 2% 18:3(n-3) without or with 0.8% 20:4(n-6) or 0.3% 22:6(n-3) from birth to 18 d. The effects of adding 20:4(n-6) or 22:6(n-3) to the formula differed among different tissues and lipids, with the brain showing resistance to change. Piglets fed formula with 20:4(n-6) had significantly higher plasma, heart and kidney phospholipid and triglyceride, and liver triglyceride 20:4(n-6), but lower plasma and tissue phospholipid 18:2(n-6) than piglets fed formula without 20:4(n-6). Supplementation with 22:6(n-3), in contrast, had no effect on plasma or tissue 18:2(n-6). Higher 22:6(n-3) in liver phospholipid (30-92% greater) and triglyceride (200% greater) in piglets fed formula with 22:6(n-3) rather than without 22:6(n-3) was accompanied by lower 20:4(n-6) in liver phosphatidylethanolamine (mean +/- SEM, 8.6 +/- 0.4 and 10.5 +/- 0.4% fatty acids, respectively), but higher 20:4(n-6) in triglyceride (5.2 +/- 0.4 and 11.5 +/- 0.5%, respectively), and higher liver, heart and kidney phospholipid 20:5(n-3). These results indicate competitive interaction between dietary 20:4(n-6) and tissue 18:2(n-6), and between dietary 20:4(n-6) and tissue 20:5(n-3), rather than 22:6(n-3). The results also show that even at low intakes, dietary 22:6(n-3) or 20:4(n-6) supplementation alters the tissue phospholipid 20:4(n-6) to 20:5(n-3) balance. Studies on the physiologic effects of dietary 20:4(n-6) and 22:6(n-3) supplementation should consider the different sensitivity among tissues to dietary fatty acids.     

Docosahexaenoic acid is transferred through maternal diet to milk and to tissues of natural milk-fed piglets

Docosahexaenoic acid (DHA) is incorporated in large amounts in structural lipids of the developing central nervous system. Milk DHA varies with maternal dietary DHA, but the effect of different intakes of DHA from milk on infant tissue fatty acids is unknown. The effect of milk high or low in DHA on the fatty acid composition of piglet brain, synaptic plasma membranes, retina, liver, plasma and RBC was studied. Pregnant sows were fed diets with 2.5 g/100 g vegetable oil until 4 d pre-partum and were then fed diets with 2.5 g/100 g soybean and canola oils or 4 g/100 g soybean oil plus 1 g/100 g fish oil to 15 d postpartum. Fish oil increased the milk DHA and eicosapentanoic acid from 0.1 to 1.5% and from 0.2 to 0.4% of fatty acids, respectively, but did not alter milk arachidonic acid. The level of DHA was significantly higher in plasma, liver and RBC phospholipids, brain and synaptic plasma membrane of 15-d-old piglets fed milk with high DHA compared with low DHA. Liver, plasma and RBC, but not brain or retina arachidonic acid, was significantly lower in piglets fed the high DHA milk compared with low DHA milk. Thus, differences in plasma, RBC and liver arachidonic acid and DHA of 15-d-old nursing piglets due to the maternal dietary fat were not accompanied by similar differences in central nervous system fatty acids. These studies show maternal DHA intake determines in part the infant plasma, RBC and liver phospholipid DHA.(ABSTRACT TRUNCATED AT 250 WORDS)     

Milk type during mixed feeding: contribution to serum cholesterol ester fatty acids in late infancy

OBJECTIVE: To evaluate the contribution of the type of milk on serum cholesterol ester fatty acids in infants receiving mixed feeding, we analyzed 3-day dietary records and serum cholesterol ester fatty acid composition of 397 seven-month-old infants. STUDY DESIGN: The infants received, in addition to solid food, only one type of milk: human milk (n = 218), a ready-to-use liquid formula (n = 139), a powdered formula (n = 33), or soy formula (n = 7). RESULTS: Mean fat intakes were low and varied from 28% to 31% of energy; the milks provided 43% to 64% of the fat. The mean polyunsaturated/saturated fat ratios of solid foods were from 0.52 to 0.63 and of milks from 0.20 to 0.45. Breast-fed infants' relative serum linoleic acid (18:2n-6) concentration was low (51.2%), whereas infants fed liquid formula had low serum oleic acid (18:1n-9) in accordance with low oleic acid content in that formula. The breast-fed infants had markedly higher serum concentrations of arachidonic acid (20:4n-6) and docosahexaenoic acid (22:6n-3) than the infants fed any of the formulas. CONCLUSION: The typical fatty acid patterns of breast- or formula-fed infants were still evident in 7-month-old infants who already received 60% to 70% of their energy from solid food. Marked differences were seen also in the relative concentrations of docosahexaenoic acid and arachidonic acid despite their small contribution in cholesterol esters.     

Polyunsaturated fatty acids in infant nutrition

The availability of long-chain polyunsaturated fatty acids (LCP), such as arachidonic (C20:4n-6) and docosahexaenoic (C22:6n-3) acids, is important for early human growth and development. The capacity for endogenous synthesis of LCP from the precursor fatty acids linoleic (C18:2n-6) and alpha-linolenic (C18:3n-3) acid is limited in preterm and probably also in term infants. In utero, LCPs seem to be transferred preferentially from the mother to the foetus by the placenta. After birth, breast-fed infants receive preformed dietary LCP with human milk. In contrast, most current infant formulae are devoid of LCP. Premature infants fed such formulae develop rapid LCP depletion of plasma and tissue lipids, which is associated with reduced visual acuity during the first postnatal months. Therefore, LCP enrichment of formulae for premature infants is desirable. Recent observations indicate that term infants fed conventional formulae also exhibit lower plasma LCP values and may show functional disadvantages, but these data require further confirmation prior to drawing definite conclusions.     

Modifying the n-3 fatty acid content of the maternal diet to determine the requirements of the fetal and suckling rat

During perinatal development, docosahexaenoic acid (22:6n-3) accumulates extensively in membrane phospholipids of the nervous system. To evaluate the n-3 fatty acid requirements of fetal and suckling rats, we investigated the accumulation of 22:6n-3 in the brain and liver of pup rats from birth to day 14 postpartum when their dams received increasing amounts of dietary 18:3n-3 (from 5 to 800 mg/100 g diet) during the pregnancy-lactation period. The fatty acid composition of brain and liver phospholipids of pups, as well as that of dam's milk, was determined. At birth, 22:6n-3 increased regularly to reach the highest level when the maternal diet contained 800 mg 18:3n-3/100 g. On days 7 and 14 postpartum, brain 22:6n-3 plateaued at a maternal dietary supply of 200 mg/100 g. Docosapentaenoic acid (22:5n-6) had the opposite temporal pattern. The unusually high concentration of eicosapentaenoic acid (20:5n-3) in liver and dam's milk observed at the highest 18:3n-3 intake suggests an excessive dietary supply of this fatty acid. All these data suggest that the n-3 fatty acid requirements of the pregnant rat are around 400 mg 18:3n-3 and those of the lactating rat at 200 mg (i.e., 0.9 and 0.45% of dietary energy, respectively). The values of 18:3n-3 and 22:6n-3 milk content which allowed brain 22:6n-3 to reach a plateau value in suckling pups were 1% of total fatty acids and 0.9% (colostrum) to 0.2% (mature milk), respectively. These levels are similar to those recommended for infant formulas.     

Net transfer and incorporation of yolk n-3 fatty acids into developing chick embryos

The effect of egg yolk fatty acid composition on the uptake and utilization of essential n-6 and n-3 fatty acids by the developing chick embryo was studied. Eggs were enriched with n-9, n-3, or n-6 fatty acids by incorporating sunflower seed high in oleic acid (C18:1 n-9), flax seed rich in linolenic acid (C18:3 n-3), or sunflower seed high in linoleic acid (C18:2 n-6) into the laying hen diets. Fertile eggs were collected and incubated. The fatty acid composition of eggs and newly hatched chicks were compared. Feeding diets containing flax seed increased (P < .05) total n-3 fatty to 528.4 mg compared with 53.9 and 39.3 mg for eggs from hens fed diets with high oleic acid or regular sunflower seed, respectively. Levels of C18:2 n-6 and monounsaturated fatty acids were higher in eggs from hens fed diets containing regular or high oleic acid sunflower seeds. Dietary fat did not influence the total lipid content of the egg yolk or total lipids of chick tissues. The fatty acid composition of the hatched progeny was significantly altered by egg yolk lipids. However, the percentage incorporation of essential n-6 and n-3 fatty acids into the progeny increased when yolk sources of these fatty acids were low. The developing chick embryo appeared to preferentially take up docosahexaenoic acid and arachidonic acid from the yolk lipids. Evidence also suggests that conversion of C18:2 n-6 and C18:2 n-3 to longer chain n-3 or n-6 fatty acids occurs during the incubation period.     

The influence of dietary fatty acid composition on N-ethyl-N-nitrosourea-induced mammary tumour incidence in the rat and on the composition of inositol- and ethanolamine-phospholipids of normal and tumour mammary tissue

This study has investigated the influence of dietary fatty acid composition on mammary tumour incidence in N-ethyl-N-nitrosourea (ENU)-treated rats and has compared the susceptibility to dietary fatty acid modification of the membrane phospholipids phosphatidylinositol (PI) and phosphatidylethanolamine (PE) from normal and tumour tissue of rat mammary gland. The incidence of mammary tumours was significantly lower in fish oil--(29%), compared with olive oil--(75%; P < 0.04) but not maize oil--(63%; P < 0.1) fed animals. No differences in PI fatty acid composition were found in normal or tumour tissue between rats fed on maize oil, olive oil or fish oil in diets from weaning. When normal and tumour tissue PI fatty acids were compared, significantly higher amounts of stearic acid (18:0) were found in tumour than normal tissue in rats given olive oil (P < 0.05). A similar trend was found in animals fed on maize oil, although differences between normal and tumour tissue did not reach a level of statistical significance (P < 0.1). In mammary PE, maize oil-fed control animals had significantly higher levels of linoleic acid (18:2n-6) than either olive oil- or fish oil-fed animals (P < 0.05, both cases) and levels of arachidonic acid were also higher in maize oil- compared with fish oil-fed animals (P < 0.05). In tumour-bearing animals no differences in PE fatty acid composition were found between the three dietary groups. When normal and tumour tissue PE fatty acids were compared, significantly lower amounts of linoleic acid (18:2n-6; P < 0.01) and significantly greater amounts of arachidonic acid (20:4n-6; P < 0.05) were found in tumour than normal tissue of rats fed on maize oil. The present study shows that the fatty acid composition of PI from both normal and tumour tissue of the mammary gland is resistant to dietary fatty acid modification. The PE fraction is more susceptible to dietary modification and in this fraction there is evidence of increased conversion of linoleic acid to arachidonic acid in tumour compared with normal tissue. Lower tumour incidence rates in rats given fish oils may in part be due to alteration in prostanoid metabolism secondary to displacement of arachidonic acid by eicosapentaenoic acid, but PE rather than PI would appear to be the most likely locus for diet-induced alteration in prostanoid synthesis in this tissue. Effects of dietary fatty acids other than on the balance of n-6 and n-3 fatty acids, and on prostanoid metabolism, should also be considered. The significance of increased stearic acid content of PI in tumours of olive oil-fed animals and the possible influence of dietary fatty acids on the capacity for stearic acid accumulation requires further study.     

Uneven distribution of desmosterol and docosahexaenoic acid in the heads and tails of monkey sperm

Previously we demonstrated high concentrations of desmosterol and docosahexaenoic acid (DHA, 22:6 n-3) in monkey testes and sperm. Desmosterol, a cholesterol precursor, is not present elsewhere in the body. High concentrations of DHA are found elsewhere only in the retina and brain. To examine the distribution of these compounds in the heads and tails of sperm, we separated them and determined their sterol, fatty acid, and phospholipid molecular species composition. Desmosterol predominated in tails (134.4 vs. 1.7 microg/10(9) cells in heads). The cholesterol content was also greater in the tails (66.2 vs. 30.3 microg/10(9) cells in heads). Sperm tails had more polyunsaturated fatty acids than the heads (34.1 vs. 12.1% of total fatty acids) which resulted mainly from the higher contents of DHA (19.6 vs. 1.1%) and arachidonic acid (20:4 n-6) (6.4 vs. 1.6%) in the tails. These differences in fatty acid composition occurred mainly in phospholipids: phosphatidyl choline and phosphatidyl ethanolamine for n-3 fatty acids and phosphatidyl serine and cardiolipin for n-6 fatty acids. Fifteen phospholipid molecular species were identified. Sperm tails had more molecular species containing unsaturated fatty acids than the heads. Our results reveal the large differences in membrane lipid composition between the heads and tails of sperm. Most (99%) of the desmosterol and DHA in sperm is located in the tail. These differences may be responsible for the different functions of these two components of sperm. The large number of double bonds in DHA, six, and in desmosterol, two, may contribute to the membrane fluidity necessary for the motility of the sperm tails.     

Effect of diet on the rate of depletion of n-3 fatty acids in the retina of the guinea pig

This study has assessed the influence of maternal n-3 long chain polyunsaturated fatty acid supply and dietary manipulation after weaning on the retinal polyunsaturated fatty acid profile. Infant guinea pigs born of dams fed one of two commercial chow diets (differing in the amount of eicosapentaenoic, docosapentaenoic, and docosahexaenoic acids) were raised in two separate experiments, and subsequently partitioned into two diet groups, one supplied with a high level of alpha-linolenic acid (canola oil supplemented), the other with a very low level of alpha-linolenic acid (safflower oil supplemented). Guinea pigs born of dams supplied with the longer chain n-3 fatty acids in the commercial pellets (experiment 2) showed higher levels of retinal docosahexaenoic acid at weaning compared with those born to dams fed chow containing only alpha-linolenic acid (experiment 1). The rate of depletion of retinal docosahexaenoic acid after weaning onto the safflower oil diet was described by a two-stage exponential decay, possibly reflecting systemic and local conservation mechanisms, in conditions of dietary n-3 fatty acid deprivation. The rate of docosahexaenoic acid depletion in the group with the lower retinal docosahexaenoic acid at weaning was more than double the rate of depletion in the group with the higher weaning docosahexaenoic acid value. The endpoint retinal docosahexaenoic acid level at 16 weeks post-weaning after dietary n-3 fatty acid depletion on the safflower oil diet in the group, which started with the lower retinal docosahexaenoic acid level, was approximately half that compared with the group from the dams fed long chain n-3 fatty acids (experiment 1, 5% (interpolated), experiment 2, 9%). These results suggest that an adequately supplied mother is capable of providing an infant with enough n-3 fatty acids to withstand a longer period of dietary deprivation imposed after weaning.     

Evaluation of the effects of omega-3 fatty acid-containing diets on the inflammatory stage of wound healing in dogs

OBJECTIVES: To ascertain the effects of dietary omega-3 (n-3) fatty acids on biochemical and histopathologic components of the inflammatory stage of wound healing. ANIMALS: 30 purpose-bred Beagles. PROCEDURE: Dogs were allotted to 5 groups of 6. Each group was fed a unique dietary fatty acid ratio of omega-6 to n-3--diet A, 5.3:1; diet B, 10.4:1; diet C, 24.1:1; diet D, 51.6:1; and diet E, 95.8:1. Dogs were fed once daily for 12 weeks, then biopsy specimens were taken from 4-day-old wounds of each dog and analyzed by gas chromatography-mass spectrometry for: prostaglandin E2 (PGE2) metabolites, and ratios of omega-6 to n-3 fatty acids, arachidonic acid (AA) to eicosapentaenoic acid (EPA), adrenic acid to docosahexaenoic acid, and PGE2 to prostaglandin E3 (PGE3) metabolites. RESULTS: Qualitative analysis was carried out on AA, EPA, adrenic acid, docosahexaenoic acid, and the major metabolite from the PGE2 and PGE3 pathway. These molecules were further quantified with respect to diet to determine significant differences. By analysis of the AA-to-EPA ratio, diet A was different from diets D and E and diets B and C were different from diet E (P < 0.05). By analysis of the PGE2-to-PGE3 metabolite ratio, diet A was different from diet E (P < 0.05). Though biochemical analysis indicated dietary dependence, histopathologic data indicated no significant difference with respect to diet groups. CONCLUSION: The biochemical component of the inflammatory stage of wound healing can be manipulated by diet. CLINICAL RELEVANCE: Omega-3 fatty acid-enriched diets can be used to control inflammation associated with dermatologic conditions.     

Isolation of the Pichia pastoris PYC1 gene encoding pyruvate carboxylase and identification of a suppressor of the pyc phenotype

It has been reported that both n-3 and n-6 octadecatrienoic acids can increase hepatic fatty acid oxidation activity. It remains unclear, however, whether different enzymes in fatty acid oxidation show a similar response to n-3 and n-6 octadecatrienoic acids. The activity of hepatic fatty acid oxidation enzymes in rats fed an oil mixture rich in alpha-linolenic acid (18:3n-3) and borage oil rich in gamma-linolenic acid (18:3n-6) was therefore compared to that in rats fed an oil mixture rich in linoleic acid (18:2n-6) and a saturated fat (palm oil) in this study. Linseed oil served as the source of 18:3n-3 for the oil mixture rich in this octadecatrienoic acid and contained 30.6% 18:3n-3 but not 18:3n-6. Borage oil contained 25.7% 18:3n-6 and 4.5% 18:3n-3. Groups of seven rats each were fed diets containing 15% various fats for 15 d. The oxidation rate of palmitoyl-CoA in the peroxisomes was higher in rats fed a fat mixture rich in 18:3n-3 (3.03 nmol/min/mg protein) and borage oil (2.89 nmol/min/mg protein) than in rats fed palm oil (2.08 nmol/min/mg protein) and a fat mixture rich in 18:2n-6 (2.15 nmol/min/mg protein). The mitochondrial palmitoyl-CoA oxidation rate was highest in rats fed a fat mixture rich in 18:3n-3 (1.93 nmol/min/mg protein), but no significant differences in this parameter were seen among the other groups (1.25-1.46 nmol/min/mg protein). Compared to palm oil and fat mixtures rich in 18:2n-6, a fat mixture rich in 18:3n-3 and borage oil significantly increased the hepatic activity of carnitine palmitoyltransferase and acyl-CoA oxidase. Compared to palm oil and a fat mixture rich in 18:2n-6, a fat mixture rich in 18:3n-3, but not fats rich in 18:3n-6, significantly decreased 3-hydroxyacyl-CoA dehydrogenase activity. Compared to palm oil and a fat mixture rich in 18:2n-6, borage oil profoundly decreased mitochondrial acyl-CoA dehydrogenase activity, but a fat mixture rich in 18:3n-3 increased it. 2,4-Dienoyl-CoA reductase activity was significantly lower in rats fed palm oil than in other groups. Compared to other fats, borage oil significantly increased delt3,delta2-enoyl-CoA isomerase activity. Activity was also significantly higher in rats fed 18:2n-6 oil than in those fed palm oil. It was confirmed that both dietary 18:3n-6 and 18:3n-3 increased fatty acid oxidation activity in the liver. These two dietary octadecatrienoic acids differ considerably, however, in how they affect individual fatty acid oxidation enzymes.     

Simultaneous measurement of [18F]FDOPA metabolism and cerebral blood flow in newborn piglets

Three diets containing either borage oil (BO) and southern hemisphere fish oil Marinol (MO), or BO and tuna orbital oil (TO), or a northern hemisphere fish oil (FO) were fed to duplicate groups of turbot (Scophthalmus maximus) of initial mean weight 1.2 g for a period of 12 weeks. The BO/MO and BO/TO diets were enriched in gamma-linolenic (18:3n-6, GLA) and eicosapentaenoic (20:5n-3, EPA) acids, and GLA and docosahexaenoic acid (22:6n-3, DHA), respectively. No differences were observed in final weights or growth rates, either between duplicate tanks or between dietary treatments. Half of the FO-fed fish sampled showed a histopathological lesion indicative of lipoid liver degeneration while the other treatments only showed a slight incidence of the same pathology. The fatty acid compositions of carcass and tissues broadly reflected the dietary input. In general, fish fed the BO/MO diet had increased levels of 18:2n-6, 18:3n-6, 20:3n-6 and 20:5n-3, but a lower level of 22:6n-3, compared to fish fed FO. In fish fed the BO/TO diet, levels of 18:2n-6, 18:3n-6, 20:3n-6 and 20:4n-6 were increased while levels of 20:5n-3 and 22:5n-3 were reduced, compared to fish fed FO. Concentrations of thromboxanes B (TXB) and leukotrienes B (LTB), derived from 20:4n-6 and 20:5n-3, were measured in plasma and stimulated blood cells. Levels of TXB2 were greatest in fish fed the BO/TO diet compared to both other treatments, while LTB4 was decreased in fish fed the BO/MO diet compared to both other treatments. In a stress test which involved anaesthesia followed by measurement of recovery times, fish fed the BO/MO diet had significantly lower recovery times compared to fish fed the FO diet.     

Effect of increasing breast milk docosahexaenoic acid on plasma and erythrocyte phospholipid fatty acids and neural indices of exclusively breast fed infants

OBJECTIVES: To determine the effect of increasing docosahexaenoic acid (DHA, 22:6 n-3) in breast milk on infant fatty acid profiles. A secondary aim was to examine aspects of neural development. DESIGN AND SETTING: Double blind, placebo controlled study of infants recruited from postnatal wards at Flinders Medical Centre. SUBJECTS: Fifty-two healthy term infants who were breast fed for at least 12 weeks and were from middle class families. INTERVENTION: Breast milk with DHA concentrations that ranged from 0.1-1.7% of total fatty acids. This was achieved by supplementation of the maternal diet for the first 12 weeks post partum. RESULTS: Breast milk with DHA was related to infant plasma (r = 0.89, P < 0.001) and erythrocyte (r = 88, P < 0.001) phospholipids in a saturable curvilinear manner so that breast milk DHA above 0.8% of total fatty acids resulted in little further increase in infant plasma or erythrocyte DHA levels. The rise in plasma and erythrocyte DHA was approximated by a fall in total n-6 polyunsaturated fatty acids. We could detect no relationship between visual evoked potential acuity (measured at 12 and 16 weeks) of infants by either the dietary grouping or the DHA status of individuals. A stepwise multiple regression showed that infant erythrocyte DHA at 12 weeks and home stimulation were the only independent factors associated with Bayley's MDI at 1 y (adjusted model r2 = 0.18, P < 0.005); while at 2 y gender and social score of the spouse were the only significant predictors of Bayley's MDI (adjusted model r2 = 0.22, P < 0.005). CONCLUSIONS: Increasing breast milk DHA levels caused a dose dependent saturable increase in infant plasma and erythrocyte phospholipid DHA. There were no long-term effects of infant DHA status on indices of neurodevelopment.     

Plasma membrane phospholipid fatty acid composition of cultured skin fibroblasts from schizophrenic patients: comparison with bipolar patients and normal subjects

Recent studies have found lower red cell plasma membrane contents and composition of the long chain polyunsaturated essential fatty acid derivatives, particularly arachidonic acid and docosahexaenoic acid, in a subgroup of chronic schizophrenic patients. These fatty acids are particularly enriched in the brain. Red blood cell levels of fatty acids are influenced by diet, medications, and other factors. Cell plasma membrane compositions of arachidonic and docosahexaenoic acids were therefore examined in cultured skin fibroblasts from 12 schizophrenic patients, 8 of whom were drug-naive and in a first episode of psychosis, 6 bipolar patients, and 8 normal control subjects. Docosahexaenoic acid as well as total n-3 essential fatty acid contents were significantly lower in cell lines from schizophrenic patients than in cell lines from bipolar patients and normal subjects, with no difference between the latter two groups. Arachidonic acid levels did not differ across the groups. The essential fatty acid profile observed is consistent with deficient delta-4 desaturase activity in schizophrenic patients.     

Fatty acid composition in maternal milk and plasma during supplementation with cod liver oil

OBJECTIVE: We investigated how cod liver oil influences the amount of essential fatty acids in mothers' breast milk. DESIGN AND INTERVENTION: Lactating mothers (n =22) were randomized into four groups 3-8 weeks after parturition. They were supplemented for 14 days with 0, 2.5, 5 and 10 ml cod liver oil (7.7 g eicosapentaenoic acid (EPA, 20:5n-3), 10.2 g docosahexaenoic acid (DHA, 22:6n-3) and 22.9 g n-3 fatty acids in total per 100 ml). RESULTS: In maternal plasma phospholipids there was an increase in the content of EPA and DHA in the group supplemented with 10 ml cod liver oil daily (P < or = 0.05). DHA concentrations in breast milk pre-supplementation ranged from 0.15 to 1.56 wt% and increased in all supplemented groups (P< or =0.05). The concentration of EPA in breast milk increased in the groups supplemented with 5 or 10 ml cod liver oil (P< or =0.05), whereas the concentration of arachidonic acid (AA, 20:4n-6) did not change in any of the supplemented groups. Total intake of DHA adjusted to body mass index (BMI), correlated to DHA concentrations in plasma (r = 0.49, P = 0.02) and breast milk (r = 0.45, P = 0.04). The concentration of tocopherol did not change during the supplementation period, neither in plasma nor in breast milk. CONCLUSION: Dietary intake of DHA is reflected in the concentration of DHA in breast milk, without affecting the concentration of AA or tocopherol.     

Regulation of the B cell response to T-dependent antigens by classical pathway complement

We observed that retinoic acid, which differentiates the human neuroblastoma SK-N-BE into mature neurons, induced an elevation in levels of polyunsaturated fatty acids, especially arachidonic acid (20:4 n-6). This effect was not induced by phorbol myristate acetate, another differentiating agent. We then explored the effects of retinoic acid on the formation of arachidonic acid and of docosahexaenoic acid from precursors and on the de novo lipid synthesis from acetate at various stages of differentiation, which was assessed by morphological (cell number and neurite outgrowth) and biochemical (protein content and thymidine incorporation) criteria. At 3 days of incubation with retinoic acid, in the n-6 series, total conversion of linoleic acid, especially to 20:3 n-6, was elevated, in association with preferential incorporation of acetate into phospholipids; in contrast, at 8 days, synthesis of 20-carbon polyunsaturated fatty acids declined, in association with enhanced incorporation in triglycerides. In the n-3 series, eicosapentaenoic acid was converted to docosahexaenoic acid in SK-N-BE, but the conversion was not affected by retinoic acid. During the early stage of neuronal differentiation, therefore, enhanced production of 20-carbon polyunsaturated fatty acids from their precursors occurred, and newly formed fatty acids were preferentially incorporated in phospholipids, possibly in association with membrane deposition. When differentiation was completed, arachidonic acid formation and incorporation of acetate in phospholipids and cholesterol declined with enhanced labeling of storage lipids.     

Effect of dietary alpha-linolenic acid and its ratio to linoleic acid on platelet and plasma fatty acids and thrombogenesis

The effect of dietary alpha-linolenic acid (18:3n-3) and its ratio to linoleic acid (18:2n-6) on platelet and plasma phospholipid (PL) fatty acid patterns and prostanoid production were studied in normolipidemic men. The study consisted of two 42-d phases. Each was divided into a 6-d pre-experimental period, during which a mixed fat diet was fed, and two-18 d experimental periods, during which a mixture of sunflower and olive oil [low 18:3n-3 content, high 18:2/18:3 ratio (LO-HI diet)], soybean oil (intermediate 18:3n-3 content, intermediate 18:2/18:3 ratio), canola oil (intermediate 18:3n-3 content, low 18:2/18:3 ratio) and a mixture of sunflower, olive and flax oil [high 18:3n-3 content, low 18:2/18:3 ratio (HI-LO diet)] provided 77% of the fat (26% of the energy) in the diet. The 18:3n-3 content and the 18:2/18:3 ratio of the experimental diets were: 0.8%, 27.4; 6.5%, 6.9; 6.6%, 3.0; and 13.4%, 2.7, respectively. There were appreciable differences in the fatty acid composition of platelet and plasma PLs. Nevertheless, 18:1n-9, 18:2n-6 and 18:3n-3 levels in PL reflected the fatty acid composition of the diets, although very little 18:3n-3 was incorporated into PL. Both the level of 18:3n-3 in the diet and the 18:2/18:3 ratio were important in influencing the levels of longer chain n-3 fatty acid, especially 20:5n-3, in platelet and plasma PL. Production of 6-keto-PGF1 alpha was significantly (P < 0.05) higher following the HI-LO diet than the LO-HI diet although dietary fat source had no effect on bleeding time or thromboxane B2 production.(ABSTRACT TRUNCATED AT 250 WORDS)     

Fatty acid composition of human brain phospholipids during normal development

The fatty acid composition of phosphatidylethanolamine (PE), ethanolamine plasmalogens (EPs), phosphatidylserine (PS), phosphatidylcholine (PC), and sphingomyelin was studied in 22 human forebrains, ranging in age from 26 prenatal weeks to 8 postnatal years. Phospholipids were separated by two-dimensional TLC, and the fatty acid methyl esters studied by capillary column GLC. Docosahexaenoic acid (22:6n-3) increased with age in PE and PC, whereas arachidonic acid (20:4n-6) remained quite constant. In EP, 22:6n-3 increased less markedly than 20:4n-6, adrenic (22:4n-6) and oleic (18:1n-9) acids being the predominant fatty acids during postnatal age. In PS, 18:1n-9 increased dramatically throughout development, and 20:4n-6 and 22:4n-6 increased only until approximately 6 months of age. Although 22:6n-3 kept quite constant during development in PS, its percentage decreased due to the accretion of other polyunsaturated fatty acids (PUFAs). As a characteristic myelin lipid, sphingomyelin was mainly constituted by very long chain saturated and monounsaturated fatty acids. Among them, nervonic acid (24:1n-9) was the major very long chain fatty acid in Sp, followed by 24:0, 26:1n-9, and 26:0, and its accretion after birth was dramatic. As myelination advanced, 18:1n-9 increased markedly in all four glycerophospholipids, predominating in EP, PS, and PC. In contrast, 22:6n-3 was the most important PUFA in PE in the mature forebrain.     

Dietary (n-3) and (n-6) polyunsaturated fatty acids rapidly modify fatty acid composition and insulin effects in rat adipocytes

The influence of dietary (n-3) compared with (n-6) polyunsatured fatty acids (PUFA) on the lipid composition and metabolism of adipocytes was evaluated in rats over a period of 1 week. Isocaloric diets comprised 16.3 g/100 g protein, 53.8 g/100 g carbohydrate and 21.4 g/100 g lipids, the latter containing either (n-3) PUFA (32.4 mol/100 mol) or (n-6) PUFA (37.8 mol/100 mol) but having identical contents of saturated, monounsaturated and total unsaturated fatty acids and identical polyunsaturated to saturated fatty acid ratios and double bond indexes. Despite comparable food intake, significantly smaller body weight increments and adipocyte size were observed in rats of the (n-3) diet group after feeding for 1 wk. Rats fed the (n-3) diet also had significantly lower concentrations of serum triglycerides, cholesterol and insulin compared with those fed the (n-6) diet, although levels of serum glucose and free fatty acids did not differ in the two dietary groups. In the (n-6) diet group, the (n-6) and (n-3) PUFA contents of plasma triglycerides, free fatty acids and phospholipids were 30-60% higher and 60-80% lower, respectively, than in the (n-3) diet group, whereas adipocyte plasma membrane phospholipids showed a significantly higher unsaturated to saturated fatty acid ratio and greater fluidity. Glycerol release in response to noradrenaline was significantly higher in the adipocytes of rats fed the (n-3) diet, whereas the antilipolytic effect of insulin generally did not differ in the two groups. Finally, insulin stimulated the transport of glucose and its incorporation into fatty acids to a lesser extent in adipocytes of (n-3) diet fed rats compared with (n-6) diet fed rats. This reduction in the metabolic effects of insulin in rats fed a (n-3) diet for 1 wk could be related to smaller numbers and a lower binding capacity of the insulin receptors on adipocytes and/or to a lesser degree of phosphorylation of the 95 kDa beta subunit of the receptor. In conclusion, dietary intake for 1 wk of (n-3) rather than (n-6) PUFA is sufficient to induce significant differences in the lipid composition and metabolic responses to insulin of rat adipocytes.     

Genetic mechanisms for duplication and multiduplication of the human CYP2D6 gene and methods for detection of duplicated CYP2D6 genes

PURPOSE: To investigate the nature and reversibility of biochemical and functional changes in the retina encountered over a single generation of dietary n-3 polyunsaturated fatty acid deficiency in guinea pigs. METHODS: Dunkin-Hartley guinea pigs were fed for 16 weeks after weaning with diets supplemented with safflower seed oil (n-3 deficient) or canola oil (n-3 sufficient, control). A number of deficient animals were repleted at 6 weeks with canola oil for 5 or 10 weeks, or at 11 weeks for 5 weeks. Electroretinograms (0.8 and 4.3 log scot td x sec) were collected at 6, 11, and 16 weeks after weaning. Conventional waveforms (a- and b-waves), oscillatory potentials, and receptoral and postreceptoral subcomponents (PIII and PII, respectively) were evaluated. Cone pathway function was assessed with 30-Hz flicker at the brighter intensity. Retinal phospholipid fatty acids were measured by capillary gas-liquid chromatography. RESULTS: Electroretinographic amplitudes showed statistically significant losses in b- and a-waves after 6 and 16 weeks of dietary n-3 deficiency, respectively. The response amplitude to 30-Hz flicker was reduced 42% after 16 weeks. Retinal docosahexaenoic acid (DHA) levels of animals maintained on the safflower oil diet for 16 weeks were 42% of levels in age-matched control subjects. There were significant losses in maximum response amplitudes (R(mPIII) and R(mPII)), although the major effect was a reduction in sensitivity of the receptoral response. Complete functional recovery was observed only in animals repleted for 10 weeks. CONCLUSIONS: Functional deficits in PIII and PII of the electroretinogram were apparent in first-generation guinea pigs fed an n-3 deficient diet. These losses showed a correlation with age and retinal DHA level, although varying degrees of dependence on the DHA level were found. All functional deficits were reversed after 10 weeks of dietary n-3 repletion. The results suggest that DHA may serve several functional and structural roles in the retina and further emphasize the requirement for DHA in the normal development of vision.