Inspiratory muscle warm-up does not improve cycling time-trial performance.

PURPOSE: This study examined the effects of an active cycling warm-up, with and without the addition of an inspiratory muscle warm-up (IMW), on 10-km cycling time-trial performance. METHODS: Ten cyclists (VO2 = 65 ± 9 mL kg(-1) min(-1)) performed a habituation 10-km cycling time-trial and three further time-trials preceded by either no warm-up (CONT), a cycling-specific warm-up (CYC) comprising three consecutive 5-min bouts at powers corresponding to 70, 80, and 90% of the gas exchange threshold, or a cycling-specific warm-up preceded by an IMW (CYC + IMW) comprising two sets of 30 inspiratory efforts against a pressure-threshold load of 40% maximal inspiratory pressure (MIP). The cycling warm-up was followed by 2-min rest before the start of the time-trial. RESULTS: Time-trial performance times during CYC (14.75 ± 0.79 min) and CYC + IMW (14.70 ± 0.75 min) were not different, although both were faster than CONT (14.99 ± 0.90 min) (P < 0.05). Throughout the time-trial, physiological (minute ventilation, breathing pattern, pulmonary gas exchange, heart rate, blood lactate concentration and pH) and perceptual (limb discomfort and dyspnoea) responses were not different between CYC and CYC + IMW. Baseline MIP during CONT and CYC was 151 ± 31 and 156 ± 39 cmH2O, respectively, and was unchanged following the time-trial. MIP increased by 8% after IMW (152 ± 27 vs. 164 ± 27 cmH2O, P < 0.05) and returned to baseline after the time-trial. CONCLUSIONS: Improvements in 10-km cycling time-trial performance following an active cycling warm-up were not magnified by the addition of an IMW. Therefore, an appropriately designed active whole-body warm-up does adequately prepare the inspiratory muscles for cycling time-trials lasting approximately 15 min.

Effects of benzyl isothiocyanate on rat and human cytochromes P450: identification of metabolites formed by P450 2B1.

Naturally occurring isothiocyanates, such as benzyl isothiocyanate (BITC), are potent and selective inhibitors of carcinogenesis induced by a variety of chemical carcinogens. These effects appear to be mediated through favorable modification of both phase I and II enzymes involved in carcinogen metabolism. The inactivation of rat and human cytochromes P450 (P450s) in microsomes and the reconstituted system by BITC was investigated. BITC is a mechanism-based inactivator of rat P450s 1A1, 1A2, 2B1, and 2E1, as well as human P450s 2B6 and 2D6. BITC was most effective in inactivating P450s 2B1, 2B6, 1A1, and 2E1, whereas the activities of human P450 2C9 and rat P450 3A2 were not altered. The concentrations required for half-maximal inactivation (K(I)) of P450s 1A1, 1A2, 2B1, and 2E1 were 35, 28, 16, and 18 microM, respectively. The corresponding values for k(inact) were 0.26, 0.09, 0.18, and 0.05 min(-1), respectively. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of P450 2B1 inactivated by [(14)C]BITC indicated specific and covalent modification of the P450 apoprotein by a metabolite of BITC. High-performance liquid chromatography analysis of the BITC metabolites revealed that benzylamine was the major metabolite and there were lesser amounts of benzoic acid, benzaldehyde, N,N'-di-benzylurea, and N,N'-di-benzylthiourea. Presumably, BITC was metabolized to the reactive benzyl isocyanate intermediate that covalently modified the P450 apoprotein or hydrolyzed to form benzylamine. BITC was an efficient inactivator of P450 2B1 with a partition ratio of approximately 11:1. This irreversible inactivation of P450s by BITC could contribute significantly to its chemopreventative action.

Comparison of hemodynamic responses to social and nonsocial stress: evaluation of an anger interview.

Hemodynamic responses to an anger interview and cognitive and physical stressors were compared, and the stability of associated hemodynamic reactions examined. Participants experienced control, handgrip, counting, and mental arithmetic tests and an anger interview on two occasions. Systolic and diastolic blood pressure, heart rate, stroke volume, and cardiac output were measured. Total peripheral resistance was also derived. The anger interview produced larger, more sustained changes in blood pressure in both sessions than the other stressors. These changes were largely a consequence of increased peripheral resistance. Consistent with previous findings, handgrip was associated with a resistance-type reaction whereas arithmetic was associated with a cardiac output-type reaction. There was low-to-modest stability of hemodynamic reactions to the interview. Further research is necessary to optimize its utility in studies of cardiovascular function. Nevertheless, the findings underscore the ability of ecologically relevant stressors to provoke unique configurations of cardiovascular activity.

Cardiovascular changes during induced emotion: an application of lang's theory of emotional imagery.

Studies of emotion have provided occasional support for physiological differentiation of affective states; however, the evidence has been inconsistent. The aims of the present study were to investigate cardiovascular changes associated with relived experiences of happiness, sadness, anger, fear, and disgust and to examine the utility of methods designed to optimize the induction of emotional responses. Thirty-four undergraduates who scored 0.5 sd above the mean on Larsen and Diener's Affect Intensity Measure described their most intense experiences of five emotions. These descriptions were then used to induce those emotions while blood pressure and other hemodynamic measures were monitored. Systolic blood pressure, diastolic blood pressure, and stroke volume differentiated among emotions. The results support the suggestion that cardiovascular activity differentiates emotional states and provide some insight into the physiological adjustments subserving such effects. The study demonstrates a method that may be applied to studies of discrete emotions.

Dietary fatty acids, membrane transport, and oxidative sensitivity in human erythrocytes.

This study examined effects of dietary n-3 fatty acids on age-related changes in erythrocyte anion transport and susceptibility to oxidation. Blood was drawn from healthy adult volunteers before and after six weeks' supplementation (nine/group) with 4.0 g/day of safflower oil (containing 2.9 g n-6 fatty acids) or fish oil (containing 1.2 g long-chain n-3 fatty acids). Following density separation of young and old erythrocytes, membrane anion transport and cell membrane lipid composition were measured. Oxidative damage was measured in erythrocyte ghosts exposed to a free radical generator. Fish oil significantly increased 16:0 and 20:5n-3 in ghosts of both young and old cells, and 22:5n-3 and 22:6n-3 in old cells alone. Safflower oil increased 16:0, 18:0, 18:1n-9, and 22:5n-6 in ghosts of young cells only. The age-dependent increase in membrane anion transport (P < 0.01) was decreased by dietary fish oil supplementation, but not by safflower oil supplementation. Safflower oil and fish oil increased the susceptibility of both young and old erythrocytes to oxidative damage by free radical generation (P < 0.001).

Dietary fatty acids and the glomerular hemodynamic response to cyclosporine in borderline hypertensive rats.

We have recently reported that cyclosporine A (CsA) decreases glomerular filtration rate in the borderline hypertensive rat (BHR), but that the glomerular filtration rate is normal when the rats are maintained on a diet supplemented with evening primrose (EP) oil. The current studies were designed to determine the glomerular hemodynamic changes responsible for this effect. A first group (PLAC-SAFF) received a diet supplemented with safflower oil (SAFF) (10% of calories) and placebo (PLAC). A second group (CsA-SAFF) received a diet supplemented with SAFF and CsA (10 mg/kg/day). A third group (CsA-EP) also received CsA, but the diet was supplemented with EP oil (10% of calories). Routine micropuncture studies were performed after five to nine weeks of treatment. Single nephron glomerular filtration rate (SNGFR) was lower in CsA-SAFF than in PLAC-SAFF (36 +/- 2 vs. 46 +/- 1 nl/min, p < 0.05). Maintenance of SNGFR in CsA-EP compared to CsA-SAFF (48 +/- 2 nl/min vs. 36 +/- 2 nl/min, P < 0.05) was due to higher values for single nephron plasma flow rate (156 +/- 16 vs. 118 +/- 9off/min, P < 0.05), and higher values for the glomerular capillary ultrafiltration coefficient (0.091 +/- 0.013 vs. 0.054 +/- 0.010 nl/s/mm Hg, P < 0.05). Since dietary fatty acids can affect prostaglandin (PG) production, we measured PGE production in isolated glomeruli. Mean values for basal production rates of PGE were greater in rats maintained on EP than in rats maintained on SAFF (3958 +/- 105 vs. 3378 +/- 146 pg PGE/mg glomerular protein, P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Comparison of three theories relating facial expressiveness to blood pressure in male and female undergraduates.

We examined differing predictions of how emotional expressions and blood pressure are related. Spontaneous positive and negative facial expressions, resting systolic blood pressure (SBP), and reactive SBP were each measured for 148 male and female undergraduates. The discharge theory of emotions proposes that few expressions will predict higher baseline SBP, and this was found for men. A mismatch theory of emotions proposes that an imbalance between positive and negative expressions will predict higher baseline SBP, and this was supported for women. Finally, coactivation theory proposes that many expressions will predict higher reactive SBP, and this was found for both men and women. These results reconcile previous conflicting findings by clarifying the conditions under which each of these theories may be supported.

Differences in the metabolism of 18:2n-6 and 18:3n-6 by the liver and kidney may explain the anti-hypertensive effect of 18:3n-6.

The present study examined the in vitro and in vivo metabolism of 18:2n-6 and 18:3n-6 by kidney and liver in the male adult spontaneously hypertensive (SHR) and normotensive (WKY) rats. In liver and kidney slices incubated for 1 h with either [1-14C]18:2n-6 or [1-14C]18:3n-6 (60 microM), substantial amounts of radioactivity were incorporated into triacylglycerol and phospholipid fractions. Approximately 15% of the radiolabeled 18:2n-6 was converted into 18:3n-6 in liver slices but no conversion was found in kidney slices. When incubated with radiolabeled 18:3n-6, over 40% of the radioactivity was metabolized mainly to 20:4n-6 in liver slices, but evenly to 20:3n-6 and 20:4n-6 in kidney slices. There were no differences between the results from SHR and those from WKY. In WKY rats given an oral bolus of radiolabeled 18:3n-6, most of the radioactivity was recovered in the liver and significantly less in the kidney. In both tissues, the radioactivity was associated initially only with 18:3n-6 and later with its elongation product, 20:3n-6. These findings indicated that the kidney, although unable to metabolize 18:2n-6, could metabolize 18:3n-6 taken up from the circulation. The effectiveness of 18:3n-6, compared to 18:2n-6, as an anti-hypertensive agent may result from the provision of a post-delta 6-desaturation metabolite which can be directly converted to blood pressure-regulating eicosanoids in the kidney.

Psychosocial stress, catecholamines, and essential fatty acid metabolism in rats.

To examine the effects of psychosocial stress and the "stress hormone," epinephrine, on essential fatty acid metabolism in rats, two studies were conducted. In the first, the effects of four weeks of (i) social isolation and (ii) group housing (control) on liver microsomal delta 6 and delta 5 n-6 desaturase activity were studied in group-reared male normotensive (Wistar Kyoto) and spontaneously hypertensive (SHR) rats (n = 5/group). The second study examined the effects of acute ip epinephrine (0.0, 1.0, 2.0, and 4.0 mg/kg) 6 hr prior to and following an ig dose (4 g/kg) of safflower oil (rich in 18:2n-6, LA) on plasma and liver LA, 20:4n-6 (AA), and LA/AA ratios in adult essential fatty acid deficient Sprague-Dawley rats (n = 6/group). In the first experiment, isolation stress significantly inhibited the activity of delta 6 (P < 0.05) and delta 5 (P < 0.01) desaturase in the normotensive rats and of delta 5 desaturase in the SHR (P < 0.05). In the second study, epinephrine increased plasma and liver LA at doses 1.0 and 2.0 mg/kg in most of the fractions examined, and suppressed AA levels. The response of the LA/AA ratio to epinephrine varied between tissues and among lipid fractions, but increased this ratio at the moderate doses (2.0-4.0 mg/kg) of epinephrine in most cases. These data suggest that psychosocial stressors are capable of inhibiting the rate limiting steps of essential fatty acid metabolism and that this response is more pronounced in the SHR than in the Wistar Kyoto. They also suggest that epinephrine is capable of altering the in vivo metabolism of essential fatty acids in the rat.

Interaction of dietary fatty acids and cyclosporine A in the borderline hypertensive rat: tissue fatty acids.

In this study we examined (i) the effects of cyclosporine A (CS) on tissue lipid composition and (ii) the effect of changes in dietary n-6 fatty acids on tissue responses to CS. Fatty acid composition of liver, kidney, heart and brain were determined after 4 wk of treatment with CS (10 mg/kg.d p.o.) in male borderline hypertensive rats (BHR, n = 4/group), whose diet was supplemented with either safflower oil or evening primrose oil (EPO). Phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine/phosphatidylinositol, triglyceride and cholesteryl ester fatty acids were measured in kidney, heart, brain and liver. The same parameters were also measured in safflower-fed BHR (n = 4) receiving placebo. The effects of CS on liver microsomal delta 9, delta 6 and delta 5 desaturases in vitro were also followed. CS affected the fatty acid composition of all tissues examined, with the greatest changes seen in the renal phosphatidylcholine and phosphatidylserine/phosphatidylinositol fractions. All CS-induced changes that occurred in the liver, brain and renal fatty acids were reversed by EPO. CS elevated delta 9 desaturase but had no effect on delta 6 and delta 5 desaturase. In light of (i) the observation that EPO normalizes renal function and blood pressure in CS-treated BHR, and (ii) the importance of the kidney in blood pressure regulation, the data suggest that the beneficial effects of EPO on CS toxicity may involve changes in renal phospholipid fatty acid profiles.

Effects of maternal dietary n-3 and n-6 fatty acids (pre- and post-delta 6 desaturation) on tissue glycerophospholipid fatty acid compositions in dams and suckling mice.

The present study examined the effects of supplementation of either 18:3n-3 or a mixture of its post-delta 6-desaturation metabolites, 20:5n-3/22:6n-3, in combination with either 18:2n-6 or its immediate delta 6-desaturation product, 18:3n-6, in the maternal diet (n-3 to n-6 ratio at 0.25) on brain, liver, heart, and kidney glycerophospholipid fatty acid composition in dams (B6D2F1 mice) and their 12-day-old suckling pups. As expected, n-3 and n-6 fatty acids competed for incorporation into tissue glycerophospholipids in both dams and their suckling pups. Feeding a 20:5n-3/22:6n-3 as compared with an 18:3n-3 rich diet increased the tissue levels of 20:5n-3 and 22:5n-3, whereas it decreased those of 20:3n-6 and 20:4n-6. Replacing 18:2n-6 with 18:3n-6 in the maternal diet increased significantly the levels of 18:3n-6, 20:3n-6, and 20:4n-6, whereas it reduced those of 20:5n-3. However, the effects of maternal dietary fats on tissue fatty acid compositions in pups were qualitatively similar to but quantitatively smaller than those in dams. The discrepancy might be due to differences in the composition of fatty acids taken up and synthesized by the dams and that transferred to the pups.

Psychological stress reverses antiaggregatory effects of dietary fish oil.

Effect of acute psychological stress on the inhibition of in vitro platelet aggregation by dietary long-chain n-3 fatty acids was studied in 20 adult males. Subjects were randomly divided into groups receiving either olive oil or fish oil (2.4 g long-chain n-3 fatty acids/day) for 4 weeks. In vitro aggregation responses to two doses of ADP collagen, and epinephrine were measured immediately prior to and following exposure to three psychological stressors (2 min each), before and after the supplementation period. Olive oil had no effect on baseline aggregatory responses, while fish oil reduced aggregatory responses to ADP and epinephrine. Exposure to the stressors had no effect upon presupplementation aggregation in either group or in the olive oil group postsupplementation. However, stress abolished antiaggregatory effects of fish oil. This reversal of the antiaggregatory effects of fish oil by mild stress suggests possible limitations of low-dose fish oil supplementation in clinical situations.

Effects of dietary fatty-acid supplementation on fatty-acid composition and deformability of young and old erythrocytes.

The effects of cell age on erythrocyte phospholipid fatty-acid composition and deformability were examined in 20 healthy adults (11 male, 9 female) prior to and following 12 weeks of dietary supplementation with 3.5 g/day of safflower oil (high in n - 6 fatty acids) or fish oil (high in n - 3 fatty acids). In the absence of dietary supplementation, old erythrocytes demonstrated an increase in filtration time (P < 0.001), an increase in membrane phospholipid total n - 6 fatty acids (P < 0.01), and a decrease in total n - 3/total n - 6 ratio (P < 0.01) compared to young erythrocytes. Both safflower and fish oil supplementation attenuated age-related differences in membrane phospholipid total n - 6 and total n - 3 fatty acids. Fish oil supplementation also increased the proportion of n - 3 fatty acids (P < 0.01) and the n - 3/n - 6 ratio (P < 0.05) in the phospholipids of both young and old erythrocytes, and eliminated age-related differences in erythrocyte filtration time by reducing the relative filtration time of the old erythrocytes.

Effect of salt-loading and spontaneous hypertension on in vitro metabolism of [1-14C]linoleic and [2-14C]dihomo-gamma-linolenic acids.

The present study compared the effect of spontaneous hypertension and salt-loading on in vitro metabolism of 18:2n-6 (linoleic acid) and 20:3n-6 (dihomo-gamma-linolenic acid). Ten weanling spontaneously hypertensive rats (SHR) and 10 normotensive Wistar-Kyoto rats (WKY) maintained on a rodent lab chow were given tap water with (n = 5) or without (n = 5) addition of 1% NaCl for 4 weeks. Thereafter, animals were killed and liver microsomes were prepared. Aliquots of microsomes suspended in the phosphate-sucrose buffer containing MgCl2, ATP, CoA, and NADPH were incubated with 0.3 microCi of [1-14C]-18:2n-6 or [2-14C]-20:3n-6 at 37 degrees C for 15 min. The activity of delta 6- and delta 5-desaturases, and the distribution of radioactivity in different lipid fractions and in phospholipid fatty acids were determined. Results showed that both spontaneous hypertension and salt-loading suppressed the desaturation of radiolabeled 18:2n-6 and of 20:3n-6. Incubation of microsomes with [1-14C]-18:2n-6 resulted in 29% of radioactivity being associated with phospholipid fraction, of which 3% was associated with 18:3n-6. Incubation with radiolabeled 20:3n-6 resulted in 30% of the radioactivity being incorporated into phospholipids, of which 50% was associated with 20:4n-6 (arachidonic acid). Salt-loading suppressed the incorporation of radiolabeled fatty acids into phospholipids, more so in SHR than in WKY. Thus, salt-loading not only suppressed the desaturation of 18:2n-6 and 20:3n-6, but also interfered with the acylation of n-6 fatty acids into the phospholipid fraction.

Effects of acute exercise on cardiovascular reactivity.

Although exercise may modulate cardiovascular reactivity to stress, its acute effects have not been studied extensively. The purpose of this study was to examine over time the acute effects of different durations of aerobic exercise on cardiovascular reactivity to stressors. Twenty-four sedentary men underwent minimal exercise, 1 or 2 hr of stationary cycling at 55% VO2max. Heart rate, blood pressure, and blood catecholamines were measured during cold pressor, Stroop, and public speech tasks 1, 3, and 24 hr after exercise. One or two hours of exercise attenuated blood pressure responses to stress. The attenuation was evident 3 hr following exercise and was most apparent on the cold pressor task. These effects were independent of epinephrine level and stress appraisal. The role of central sympathetic processes in the effects of exercise and methodologic implications are discussed.

Effects of repeated gestation and lactation on milk n-6 fatty acid composition in rats fed on a diet rich in 18:2n-6 or 18:3n-6.

The present study examined the effect of repeated gestation and lactation on the levels of long-chain n-6 polyunsaturated fatty acids in rat milk fat, and examined whether such levels might be modulated by supplementing the diet of the lactating dams with either (g/kg) 50 safflower oil (SFO; containing 800 g 18:2n-6/kg), or 50 evening primrose oil (EPO; containing 720 g 18:2n-6 and 90 g 18:3n-6/kg). The milk was collected at three different times (days 1, 8 and 15) in each given lactation period from female Sprague-Dawley rats which were successively bred for four pregnancies and lactations. Results showed that dietary fat and breeding frequency had no significant effects on milk triacylglycerol content, but they modified the pattern of milk fatty acids in both triacylglycerol and phospholipid fractions. After three or four successive breedings rats fed on EPO produced milk containing less saturated but more monounsaturated and polyunsaturated fatty acids compared with those fed on SFO. During the course of lactation the levels of n-6 metabolites, e.g. 18:3n-6, 20:3n-6 and 20:4n-6, in milk fat declined progressively. However, they were consistently higher in the EPO group than in the SFO group. These findings suggest that the levels of long-chain n-6 metabolites in the milk fat may be increased through supplementing the maternal diet with 18:3n-6.

Attenuation of cyclosporine-induced hypertension by dietary fatty acids in the borderline hypertensive rat.

The effects of dietary (10% calories) safflower (SAF), evening primrose (EPO), and fish oil (F) as sources of linoleic acid (control), gamma-linolenic acid, and long-chain n-3 fatty acids, respectively, on cardiovascular and renal responses to chronic (5 weeks) cyclosporine administration were studied in male borderline hypertensive rats. In one experiment (n = 9/group), oral administration of CsA at 0.1 mg/kg.day significantly increased awake systolic blood pressure vs. placebo in SAF-fed rats (P less than 0.01). This increase was prevented by both EPO (P less than 0.001) and F (P less than 0.01), in the absence of group differences in body weight gain or plasma electrolyte levels. In a second experiment, CsA also increased blood pressure vs. placebo in SAF-fed rats (P less than 0.001). While this increase was prevented by EPO (P less than 0.001), F had no significant effect. Differences in group blood pressure responses were not explained by group differences in body weight gain or trough levels of blood CsA. Renal function, assessed in anesthetized rats after week 5, demonstrated a CsA-related (10 mg/kg.day) decrease in whole-kidney GFR in SAF-fed animals vs. placebo (P less than 0.05) that was prevented by EPO and attenuated by F. EPO and F also tended to reduce the CsA-induced elevation in renovascular resistance, but this difference did not reach statistical significance. These findings suggest the potential of dietary EPO and F to offset nephrotoxic effects of CsA administration, and suggest that EPO may also be useful in countering CsA-induced hypertension.

Effect of maternal dietary fats with variable n-3/n-6 ratios on tissue fatty acid composition in suckling mice.

This report examines the distribution of n-3 and n-6 fatty acids in heart, kidney and liver phosphatidylcholine and phosphatidylethanolamine of suckling mice from dams fed a fat-supplemented diet with variable n-3/n-6 ratios. After conception and throughout the pregnancy and lactation period, dams were fed a fat-free liquid diet supplemented with 20% by energy of oil mixtures (fish oil concentrate, rich in 20:5n-3 and 22:6n-3, and safflower oil concentrate, rich in 18:2n-6). The diets contained similar amounts of combined n-3 and n-6 fatty acids but variable ratios of n-3 to n-6 fatty acids (0, 0.25, 0.5, 1, 2 and 4). In 12-day-old suckling mice, as the n-3/n-6 ratio in the maternal diet increased (up to approx. 0.5), the tissue levels of 20:5n-3, 22:5n-3 and 22:6n-3 increased, whereas those of 18:2n-6 and 20:4n-6 decreased. The responses were similar in both phospholipid subclasses, but varied between different tissues. Generally, the n-3/n-6 ratios were significantly greater in pup tissues than in milk fat, indicating preferential incorporation of n-3 over n-6 fatty acids into phospholipids during growth. However, the incorporation of n-3 fatty acids in pups was significantly suppressed whereas that of n-6 fatty acids was increased when 18:2n-6 was replaced by its delta 6-desaturation product, 18:3n-6 (concentrated from evening primrose oil), as the source of n-6 fatty acid.(ABSTRACT TRUNCATED AT 250 WORDS)

The effects of dietary n-3/n-6 ratio on brain development in the mouse: a dose response study with long-chain n-3 fatty acids.

This study examines the effects of the ratio of n-3/n-6 fatty acids (FA) on brain development in mice when long-chain n-3 FA are supplied in the diet. From conception until 12 days after birth, B6D2F1 mice were fed liquid diets, each providing 10% of energy from olive oil, and a further 10% from different combinations of free FA concentrates derived from safflower oil (18:2n-6), and fish oil (20:5n-3 and 22:6n-3). The range of dietary n-3/n-6 ratios was 0, 0.25, 0.5, 1.0, 2.0 and 4.0, with an n-6 content of greater than 1.5% of energy in all diets, and similar levels of total polyunsaturated fatty acids (PUFA). In an additional group of ratio 0.5, 18:2n-6 was partially replaced by its delta 6 desaturation product, 18:3n-6. Biochemical analyses were conducted on 12-day-old pup brains, as well as on samples of maternal milk. No obvious effects on overall pup growth and development were observed, apart from a smaller litter size at ratio 1. Co-variance analysis indicated that increasing the n-3/n-6 ratio was associated with slightly smaller brains, relative to body weight. We found that 18:2n-6 and 20:5n-3 were the predominant n-6 and n-3 FA in the milk; in the brain these were 20:4n-6 and 22:6n-3, respectively. Increasing dietary n-3/n-6 ratios generally resulted in an increase in n-3 FA, with a corresponding decrease in n-6 FA.(ABSTRACT TRUNCATED AT 250 WORDS)