Combined crystal structure prediction and high-pressure crystallization in rational pharmaceutical polymorph screening.

Organic molecules, such as pharmaceuticals, agro-chemicals and pigments, frequently form several crystal polymorphs with different physicochemical properties. Finding polymorphs has long been a purely experimental game of trial-and-error. Here we utilize in silico polymorph screening in combination with rationally planned crystallization experiments to study the polymorphism of the pharmaceutical compound Dalcetrapib, with 10 torsional degrees of freedom one of the most flexible molecules ever studied computationally. The experimental crystal polymorphs are found at the bottom of the calculated lattice energy landscape, and two predicted structures are identified as candidates for a missing, thermodynamically more stable polymorph. Pressure-dependent stability calculations suggested high pressure as a means to bring these polymorphs into existence. Subsequently, one of them could indeed be crystallized in the 0.02 to 0.50 GPa pressure range and was found to be metastable at ambient pressure, effectively derisking the appearance of a more stable polymorph during late-stage development of Dalcetrapib.

Docosahexaenoic acid synthesis from alpha-linolenic acid is inhibited by diets high in polyunsaturated fatty acids.

The conversion of the plant-derived omega-3 (n-3) α-linolenic acid (ALA, 18:3n-3) to the long-chain eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) can be increased by ALA sufficient diets compared to ALA deficient diets. Diets containing ALA above an optimal level result in no further increase in DHA levels in animals and humans. The present study evaluates means of maximizing plasma DHA accumulation by systematically varying both linoleic acid (LA, 18:2n-6) and ALA dietary level. Weanling rats were fed one of 54 diets for three weeks. The diets varied in the percentage of energy (en%) of LA (0.07-17.1 en%) and ALA (0.02-12.1 en%) by manipulating both the fat content and the balance of vegetable oils. The peak of plasma phospholipid DHA (>8% total fatty acids) was attained as a result of feeding a narrow dietary range of 1-3 en% ALA and 1-2 en% LA but was suppressed to basal levels (∼2% total fatty acids) at dietary intakes of total polyunsaturated fatty acids (PUFA) above 3 en%. We conclude it is possible to enhance the DHA status of rats fed diets containing ALA as the only source of n-3 fatty acids but only when the level of dietary PUFA is low (<3 en%).

Rotation-libration and rotor-rotor coupling in 4-methylpyridine.

The low temperature rotational dynamics of methyl groups in 4-methylpyridine is analyzed in terms of a model potential including rotation-libration and rotor-rotor coupling. The parameters of the model potential are adjusted by comparison of calculated with published and newly recorded inelastic neutron scattering spectra. Initial evaluations of the potential parameters of the model are obtained from molecular mechanics calculations. Experimental spectra are calculated from these potentials by numerical solution of Schrödinger's equation for clusters of coupled rotors embedded in a bigger ensemble of rotors treated in the mean field approximation. Adjustment of the potential parameters leads to excellent agreement with the experimental spectra of protonated 4-methylpyridine, measured at well-defined spin temperatures. At higher levels of deuteration, agreement with experiment is qualitative, only. The observed deviations are attributed to the increasing frustration of the system of coupled methyl groups and mutual localization, effects leading to a phase transition around 5.5 K in isotopic mixtures, as shown in diffraction experiments.

Perinatal characteristics may influence the outcome of visual acuity.

Visual-evoked potential (VEP) acuity has been used to assess the effects of dietary fats on the integrity of the visual pathway of infants. We investigated prognostic determinants of VEP acuity at 16 wk of age. The results of two randomized dietary intervention trials designed to assess the effect of dietary fatty acids on the visual development of term infants were combined. At entry to both trials (approximately day 5 of life), a blood sample to assess polyunsaturated fatty acid (PUFA) status was collected along with sociodemographic and perinatal characteristics. At 16 +/- 0.9 wk of age, infants underwent VEP testing to measure acuity. There was no effect of dietary treatment on these outcomes within or between trials. Multiple linear regression models were constructed to investigate the effect of perinatal and nutritional variables at study entry on VEP acuity of 185 infants. Higher birth weight was associated with an ability to resolve smaller checkerboard patterns [r2 = 0.05; 95% confidence interval (Cl), -0.10, -0.04 log units]. Male gender (r2 = 0.03; 95% Cl, 0.01, 0.07 log units), day 5 plasma 22:5n-6 (r2 = 0.04; 95% Cl, 0.02, 0.20 log units), day 5 red cell membrane 20:3n-9 (r2 = 0.03; 95% Cl, 0.03, 0.13 log units), and the number of smokers in the household (r2 = 0.02; 95% Cl, 0.00, 0.04 log units) were all associated with poorer VEP acuity scores. It is possible that a combination of perinatal factors could accumulate to either mask or enhance effects of diet on VEP acuity, given the relatively modest effects of long-chain PUFA on visual outcome.

Transforming growth factor beta in human milk does not change in response to modest intakes of docosahexaenoic acid.

Long-chain polyunsaturated fatty acids have been associated with aspects of immune regulation including cytokine production. The purpose of this study was to investigate the effect of maternal dietary supplementation with tuna oil, rich in docosahexaenoic acid (DHA), on the concentration of transforming growth factor beta 1 (TGFbeta1) and TGFbeta2 in breast milk. In this randomized, dietary intervention trial, mothers of term infants consumed a daily supplement of 2000 mg oil containing either placebo (n = 40), 300 mg DHA (n = 40), or 600 mg DHA (n = 40). The DHA increase in milk and plasma was proportional to dietary DHA. There was no relationship between milk DHA status and TGFbeta1 and TGFbeta2 levels.

Biochemical effects of a diet containing foods enriched with n-3 fatty acids.

BACKGROUND: Results of many studies indicate that consumption of n-3 fatty acids can benefit persons with cardiovascular disease and rheumatoid arthritis. However, encapsulated fish oil is unlikely to be suited to lifetime daily use and recommendations to increase fish intake have not been effective. OBJECTIVE: The objective was to examine the effectiveness of a diet that incorporates foods rich in n-3 fatty acids in elevating tissue concentrations of eicosapentaenoic acid and in suppressing the production of inflammatory mediators. DESIGN: Healthy male volunteers were provided with foods that were enriched in alpha-linolenic acid (cooking oil, margarine, salad dressing, and mayonnaise) and eicosapentaenoic and docosahexaenoic acids (sausages and savory dip) and with foods naturally rich in n-3 fatty acids, such as flaxseed meal and fish. Subjects incorporated these products into their food at home for 4 wk. Fatty acid intakes, cellular and plasma fatty acid concentrations, and monocyte-derived eicosanoid and cytokine production were measured. RESULTS: Analyses of dietary records indicated that intake of eicosapentaenoic acid plus docosahexaenoic acid averaged 1.8 g/d and intake of alpha-linolenic acid averaged 9. 0 g/d. These intakes led to an average 3-fold increase in eicosapentaenoic acid in plasma, platelet, and mononuclear cell phospholipids. Thromboxane B(2), prostaglandin E(2), and interleukin 1beta synthesis decreased by 36%, 26%, and 20% (P < 0.05), respectively. CONCLUSIONS: Foods that are strategically or naturally enriched in n-3 fatty acids can be used to achieve desired biochemical effects without the ingestion of supplements or a change in dietary habits. A wide range of n-3-enriched foods could be developed to support large-scale programs on the basis of the therapeutic and disease-preventive effects of n-3 fatty acids.

A critical appraisal of the role of dietary long-chain polyunsaturated fatty acids on neural indices of term infants: a randomized, controlled trial.

OBJECTIVE: To determine whether dietary long-chain polyunsaturated fatty acids (LCPUFA), such as docosahexaenoic acid (DHA) and arachidonic acid, affect visual evoked potential (VEP) acuity of formula-fed infants, relative to breastfed infants. A secondary objective was to assess the effect of LCPUFA on Bayley's mental developmental index (MDI) and psychomotor developmental index (PDI). METHODS: Formula-fed infants were randomly allocated, in a double-blind manner, to either a placebo (no LCPUFA; n = 21), DHA supplemented (n = 23), or DHA+arachidonic acid supplemented formula (n = 24). Infants were fed their assigned formula from the first week of life until 1 year of age. A parallel reference group of breastfed infants was recruited and followed (n = 46). Infant VEP acuity was assessed at 16 and 34 weeks, and Bayley's MDI and PDI were assessed at 1 and 2 years of age. RESULTS: There were no differences among the randomized formula groups for VEP acuity at either 16 or 34 weeks of age. Breastfed infants had better VEP acuity at 34 weeks of age, but not at 16 weeks, compared with all formula-fed infants. Bayley's MDI and PDI were similar in the 3 formula-fed groups at 1 and 2 years. Breastfed infants had higher MDI scores than formula-fed infants at 2 years of age even after adjusting for environmental variables. CONCLUSIONS: LCPUFA supplementation did not influence VEP acuity development in these well-nourished, formula-fed infants.

A randomized trial of different ratios of linoleic to alpha-linolenic acid in the diet of term infants: effects on visual function and growth.

BACKGROUND: There are nutritional recommendations that the ratio of linoleic to alpha-linolenic acid (LA:ALA) in formula for term infants be between 5:1 and 15:1. These recommendations were made in the absence of data on functional or clinical outcomes. OBJECTIVE: We compared the fatty acid status, visual evoked potential (VEP) acuity, and growth of term infants fed formula containing an LA:ALA of 10:1 or 5:1 with those of a breast-fed reference cohort. DESIGN: Formula-fed infants were allocated randomly in a double-blind fashion to receive formula with an LA:ALA of either 10:1 (16.9:1.7; n = 36) or 5:1 (16.3:3.3; n = 37) from near birth to 34 wk of age. Increased ALA was attained by replacing soy oil with low-erucic acid cannola oil. A parallel group of breast-fed infants was also recruited. Infant growth and fatty acid status were assessed at 6, 16, and 34 wk of age. VEP acuity was assessed at 16 and 34 wk. RESULTS: Infants fed the 5:1 formula had greater docosahexaenoic acid (DHA) concentrations in plasma and erythrocyte phospholipids than did infants fed the 10:1 formula, but DHA concentrations of infants fed the 5:1 formula remained less than those in breast-fed infants. The VEP acuity of all formula-fed and breast-fed infants was not significantly different at 16 and 34 wk of age. At birth, infants fed the 5:1 formula were heavier, were longer, and had a greater head circumference than infants assigned to the 10:1 formula group; this differential was maintained throughout the trial. The rate of gain in weight, length, and head circumference was not significantly different between the 2 formula-fed groups, although breast-fed infants had lower weight and length gains than did formula-fed infants between 16 and 34 wk of age. CONCLUSION: Lowering the LA:ALA in formula from 10:1 to 5:1 by using low-erucic acid canola oil resulted in a modest increase in plasma DHA but had no effect on VEP acuity or growth rate.

Dietary long-chain polyunsaturated fatty acids do not influence growth of term infants: A randomized clinical trial.

OBJECTIVE: To determine if dietary long-chain polyunsaturated fatty acids (LCPUFA) affect the growth of formula-fed infants relative to breastfed infants. METHODS: A randomized, double-blind trial of three formula-fed groups and a parallel reference group of breastfed infants was conducted. The three treatments were a placebo (no LCPUFA), docosahexaenoic acid (DHA) supplemented, and DHA plus arachidonic acid supplemented formulas fed for 12 months. Infant weight, length, head circumference, and fatty acid status were assessed at 6, 16, 34 weeks, and 1 year of age. Anthropometrics were repeated at 2 years of age. RESULTS: There were no differences between the randomized formula groups for weight, length, or head circumference even after adjustment for gender, gestational age, and the actual age at assessment. Post hoc regressions demonstrated a small negative association between DHA status at 16 weeks of age and weight at 1 and 2 years. CONCLUSIONS: Dietary LCPUFA do not influence growth of healthy term infants to a clinically significant degree.

The elimination of sodium and potassium hydroxides from desiccated soda lime diminishes degradation of desflurane to carbon monoxide and sevoflurane to compound A but does not compromise carbon dioxide absorption.

UNLABELLED: Normal (hydrated) soda lime absorbent (approximately 95% calcium hydroxide [Ca(OH)2], the remaining 5% consisting of a mixture of sodium hydroxide [NaOH] and potassium hydroxide [KOH]) degrades sevoflurane to the nephrotoxin Compound A, and desiccated soda lime degrades desflurane, enflurane, and isoflurane to carbon monoxide (CO). We examined whether the bases in soda lime differed in their capacities to contribute to the production of these toxic substances by degradation of the inhaled anesthetics. Our results indicate that NaOH and KOH are the primary determinants of degradation of desflurane to CO and modestly augment production of Compound A from sevoflurane. Elimination of these bases decreases CO production 10-fold and decreases average inspired Compound A by up to 41%. These salutary effects can be achieved with only slight decreases in the capacity of the remaining Ca(OH)2 to absorb carbon dioxide. IMPLICATIONS: The soda lime bases used to absorb carbon dioxide from anesthetic circuits can degrade inhaled anesthetics to compounds such as carbon monoxide and the nephrotoxin, Compound A. Elimination of the bases sodium hydroxide and potassium hydroxide decreases production of these noxious compounds without materially decreasing the capacity of the remaining base, Ca(OH)2, to absorb carbon dioxide.

The effect of breast feeding on lymphocyte subpopulations in healthy term infants at 6 months of age.

Breast milk contains many immunologically active components that influence the development of the immune system of the breast-fed infant. The purpose of this study was to investigate the difference in specific lymphocyte subsets between breast-fed and formula-fed 6-mo-old infants. Peripheral blood samples were collected from 79 breast-fed (< 120 mL formula/wk) and 69 formula-fed (breast-fed < 4 wk) infants at 6 mo. All infants had been born at term and had no known illness at the time of blood collection. Packed cells from whole blood were incubated with fluorochrome-labeled monoclonal antibodies, followed by erythrocyte lysis. Washed lymphocytes were analyzed by two-color direct immunofluorescence on a flow cytometer. The percentage of T and B lymphocytes in the peripheral blood of 6-mo-old infants was the same, regardless of feeding regimen. However, the relative frequency of natural killer (NK) cells was greater in breast-fed infants than in formula-fed infants (9.7% vs 7.1%; p < 0.001). The percentage of cells expressing CD4 was lower in breast-fed infants than in formula-fed infants (47.3% vs 50.9%; p < 0.005), and that of cells expressing CD8 was greater (18.0% vs 16.4%; p < 0.05). As a result, the CD4:CD8 ratio in breast-fed infants was lower than that in formula-fed infants (2.8 vs 3.3; p < 0.005). The absolute size of the lymphocyte subpopulations T, B, and CD8+ was the same for each of the two populations of infants. However, breast-fed infants had fewer CD4+ T cells (p < 0.05) and a greater number of NK cells (p < 0.01) than the age-matched formula-fed infants. The immunophenotypic differences between breast-fed and formula-fed infants are consistent with reported age-related changes, suggesting greater maturity in the development of the immune system of breast-fed infants.

Changing from isoflurane to desflurane toward the end of anesthesia does not accelerate recovery in humans.

BACKGROUND: In an attempt to combine the advantage of the lower solubilities of new inhaled anesthetics with the lesser cost of older anesthetics, some clinicians substitute the former for the latter toward the end of anesthesia. The authors tried to determine whether substituting desflurane for isoflurane in the last 30 min of a 120-min anesthetic would accelerate recovery. METHODS: Five volunteers were anesthetized three times for 2 h using a fresh gas inflow of 2 l/min: 1.25 minimum alveolar concentration (MAC) desflurane, 1.25 MAC isoflurane, and 1.25 MAC isoflurane for 90 min followed by 30 min of desflurane concentrations sufficient to achieve a total of 1.25 MAC equivalent ("crossover"). Recovery from anesthesia was assessed by the time to respond to commands, by orientation, and by tests of cognitive function. RESULTS: Compared with isoflurane, the crossover technique did not accelerate early or late recovery (P > 0.05). Recovery from isoflurane or the crossover anesthetic was significantly longer than after desflurane (P < 0.05). Times to response to commands for isoflurane, the crossover anesthetic, and desflurane were 23 +/- 5 min (mean +/- SD), 21 +/- 5 min, and 11 +/- 1 min, respectively, and to orientation the times were 27 +/- 7 min, 25 +/- 5 min, and 13 +/- 2 min, respectively. Cognitive test performance returned to reference values 15-30 min sooner after desflurane than after isoflurane or the crossover anesthetic. Isoflurane cognitive test performance did not differ from that with the crossover anesthetic at any time. CONCLUSIONS: Substituting desflurane for isoflurane during the latter part of anesthesia does not improve recovery, in part because partial rebreathing through a semiclosed circuit limits elimination of isoflurane during the crossover period. Although higher fresh gas flow during the crossover period would speed isoflurane elimination, the amount of desflurane used and, therefore, the cost would increase.

In rats breathing from a nonrebreathing system, substitution of desflurane for isoflurane toward the end of anesthesia incompletely restores the time of recovery toward that of desflurane.

UNLABELLED: The lower solubility of desflurane allows a more rapid emergence from anesthesia than after anesthesia with the more soluble but less expensive anesthetic, isoflurane. Some practitioners use isoflurane for maintenance of anesthesia, crossing over to desflurane later in maintenance in an attempt to combine the cost-effectiveness of isoflurane with the rapid emergence from desflurane. We hypothesized that this maneuver would not accomplish its goals. Twenty-four male Sprague-Dawley rats received 1.2 minimum alveolar anesthetic concentration (MAC) of desflurane for the final 15, 30, or 60 min of a 2-h, 1.2-MAC isoflurane anesthetic in a nonrebreathing anesthesia system. We measured the time from cessation of anesthetic administration to the time each rat righted himself twice. Immediately after righting for the second time, we tested each rat's ability to remain atop a rotating rod (Rota-Rod) for 60 s continuously. Early (righting reflex) and late (Rota-Rod) recovery occurred more rapidly (P < 0.001) after 120 min of anesthesia with desflurane alone than after 120 min of anesthesia with isoflurane alone. A cross-over period of 30 min or longer produced a righting reflex time that did not differ from that found with desflurane alone, but a 15-min cross-over did not. Progressively longer cross-over periods led to proportionally better Rota-Rod performance, but no cross-over duration produced the rapidity of recovery seen with desflurane alone. We concluded that in a nonrebreathing system, switching to desflurane during the last 30 min of anesthesia substantially improved early recovery but produced a much smaller improvement in later recovery. IMPLICATIONS: The newer inhaled anesthetics offer the advantage of lower solubility, and thus more rapid emergence from anesthesia, than do the older inhaled anesthetics. However, they can be more expensive to use. This study demonstrates that substituting the newer anesthetic, desflurane, toward the end of anesthesia for an older anesthetic of greater solubility, isoflurane, does not produce recovery comparable to that of desflurane alone. Furthermore, this technique can be more costly than using desflurane throughout anesthesia.

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.

Effect of dietary n-9 eicosatrienoic acid on the fatty acid composition of plasma lipid fractions and tissue phospholipids.

n-9 Eicosatrienoic acid (ETrA), also known as Mead acid, is a minor fatty acid in essential fatty acid (EFA)-sufficient healthy subjects but is found at increased levels in EFA deficiency. This study examined the influence of dietary ETrA from a biological source on plasma and tissue ETrA. A synthetic fat-free diet was prepared to which was added Mut 48 oil which contains 19% ETrA (wt%) as well as other n-9 fatty acids. Blends of vegetable oils were used to achieve overall diets with 5% fat (wt%) and varying amounts of ETrA at two different dietary levels of linoleic acid (LA), approximately 4.4 and 19% of total fatty acids. These diets were fed to 5-week-old Dark Agouti rats for four weeks. Plasma lipid fractions and liver, spleen, and peritoneal exudate (PE) cells were analyzed for fatty acid composition. ETrA was present at up to 20% total fatty acids in plasma triglyceride, cholesterol ester, and phospholipid fractions. ETrA also accumulated to substantial levels in phospholipids of liver and spleen (up to 15% of total fatty acids) and PE cells (up to 11%). ETrA was found in plasma and tissue phospholipids in proportion to the amount of ETrA present in the diet. The incorporation was reduced in diets with higher LA content compared to diets containing similar amounts of ETrA but lower LA. All rats remained apparently healthy, and histological survey of major organs revealed no abnormality. While the long-term implications for health of ingestion of diets rich in ETrA remain to be established, rats appear to tolerate high levels of dietary ETrA without adverse effects. Dietary enrichment with ETrA warrants further investigation for possible beneficial effects in models of inflammation and autoimmunity, as well as in other conditions in which mediators derived from n-6 fatty acids can affect homeostasis adversely.

Dietary (n-9) eicosatrienoic acid from a cultured fungus inhibits leukotriene B4 synthesis in rats and the effect is modified by dietary linoleic acid.

Eicosatrienoic acid (ETrA) is the (n-9) homologue of (n-6) arachidonic acid (AA) and (n-3) eicosapentaenoic acid (EPA). ETrA can be synthesized endogeneously, but tissue levels are normally undetectable except in essential fatty acid (EFA) deficiency. An ETrA-rich oil extracted from a cultured fungus was used to prepare diets which had varying levels of ETrA (0-8 g/kg diet) in combination with one of two levels of linoleic acid (LA, 2.2 or 9.5 g/kg diet). All diets were sufficient in essential fatty acids. Groups of rats were fed these diets for 4 wk after which leucocyte fatty acid content and leukotriene B4 (LTB4) synthesis were measured. The influence of dietary LA on ETrA accumulation in cells was studied and correlations with LTB4 synthesis determined. ETrA was efficiently incorporated into peritoneal exudate cell (PEC) phospholipids with no evident saturation being observed with levels up to 10 mol/100 mol total fatty acids in peritoneal exudate cells. Cellular ETrA levels were lower (P < 0.001) in rats fed the higher level of LA. ETrA accumulation in peritoneal exudate cells correlated (r(2) = 0.63, P < 0.05) with reduced LTB4 synthesis which was attributable to LTA hydrolase inhibition. Thus, dietary ETrA from a biological source can accumulate in leucocytes and suppress inflammatory eicosanoid synthesis. The findings justify further studies into the biochemical and anti-inflammatory effects of dietary ETrA, which could be incorporated into palatable food additives.

Effect of maternal docosahexaenoic acid (DHA) supplementation on breast milk composition.

OBJECTIVE: To assess the effect of varying maternal intake of docosahexaenoic acid (DHA, 22 : 6n-3), in the absence of other dietary polyunsaturates, on breast milk fatty acids. DESIGN AND INTERVENTION: Lactating mothers were randomised on day 5 post-partum to groups consuming equal numbers of capsules but containing either placebo or an oil containing DHA (43%) as its only polyunsaturate to receive 0, 0.2, 0.4, 0.9, 1.3 g DHA/day. Breast milk fatty acids as well as maternal plasma and erythrocyte phospholipids were assessed at 12 weeks post partum by capillary gas chromatography. RESULTS: Breast milk DHA levels ranged from 0.2 to 1.7% of total fatty acids and increased in a dose dependent manner (r2 = 0.89, P < 0.01). Maternal plasma (r2 = 0.71, P < 0.01) and erythrocyte (r2 = 0.77, P < 0.01) phospholipid DHA levels increased and were also strongly associated with dietary dose of DHA. Increasing maternal dietary doses of DHA did not affect breast milk arachidonic acid (AA, 20 : 4n-6) levels or antioxidant status as measured by plasma vitamin A or E levels. CONCLUSIONS: Our results have demonstrated that DHA in the diet has a strong, specific and dose-dependent effect on breast milk DHA.

Effect of dietary docosahexaenoic acid on brain composition and neural function in term infants.

There is a need to determine whether there is a dietary requirement for docosahexaenoic acid (DHA, 22:6n-3) by term infants to achieve their full developmental potential. Studies of brain fatty acid composition demonstrated that infants who were breast fed had greater levels of cerebral cortex DHA than did infants who were formula fed, suggesting that DHA in the cerebrum is dependent on a supply in the diet. Some physiological studies reported that electrophysiological and behavioral assessments of visual function were improved in breast-fed infants relative to those fed formula and that this was related to the length of breast feeding. While some randomized studies of DHA supplementation of infant formula to term infants demonstrated that the visual function of formula-fed infants could be improved to breast-fed levels by adding DHA to formula, others failed to demonstrate an effect. Variations in dietary treatments and methods of assessment make comparison of the studies difficult. Further work is necessary to rigorously establish if there are long-term benefits of dietary DHA to the term infant.

Is dietary docosahexaenoic acid essential for term infants?

There is a need to determine whether there is a dietary requirement for docosahexaenoic acid (DHA, 22:6 omega 3) by term infants to achieve their full developmental potential. Studies of brain fatty acid composition have demonstrated that infants who were breast fed have greater levels of cerebral cortex DHA than infants who were formula fed, suggesting that DHA in the cerebrum is dependent on a supply in the diet. Some physiological studies report that electrophysiological and behavioral assessments of visual function are improved in breast-fed infants relative to those fed formula, and that this is related to the level of DHA in their erythrocytes, whereas other studies demonstrate equivalent visual function between breast- and formula-fed infants. However, randomized studies of DHA supplementation of infant formula demonstrate that the visual function of formula-fed infants can be improved to breast-fed levels by adding DHA to formula. Further work is necessary to establish if there are long-term benefits of dietary DHA to the term infant.

Erythrocyte fatty acids of term infants fed either breast milk, standard formula, or formula supplemented with long-chain polyunsaturates.

The purpose of our study was to assess whether a supplement of fish oil (FO) and evening primrose oil (EPO) for formula-fed infants was capable of avoiding reductions in erythrocyte docosahexaenoic acid (DHA, 22:6n-3) and arachidonic acid (AA, 20:4n-6) associated with standard formula feeding. Healthy, term infants, whose mothers chose to formula feed, were randomized to either a placebo or supplemented formula for their first 30 wk of life. A reference group of beast-fed infants also was enrolled. Erythrocyte fatty acids were measured by capillary gas chromatography on day 5 and in weeks 6, 16, and 30. Supplementation of formula with 0.36% of total fatty acids as DHA resulted in erythrocyte DHA being maintained at or above breast-fed levels for the entire 30-wk study period, and breast feeding (0.21% DHA) resulted in a modest fall in erythrocyte DHA relative to baseline (day 5) values. The level of erythrocyte DHA in placebo formula-fed infants was halved by week 16. AA levels decreased in all infants in the first six weeks, but the levels in breast- and placebo formula-fed infants increased with age and returned to approximate baseline (day 5) values by 16 and 30 wk of age, respectively. Erythrocyte AA in FO+EPO-supplemented infants remained low and below breast- and placebo formula-fed levels. Our data suggest that dietary supplementation with DHA at 0.36% total fatty acids results in erythrocyte DHA levels above those found in breast-fed infants. EPO supplementation was not effective at maintaining erythrocyte AA when given with FO.