Comparison of muscle fatty acid profiles and cholesterol concentrations of bison, beef cattle, elk, and chicken.

The objective of this study was to compare fatty acid weight percentages and cholesterol concentrations of longissimus dorsi (LD), semitendinosus (ST), and supraspinatus (SS) muscles (n = 10 for each) of range bison (31 mo of age), feedlot-finished bison (18 mo of age), range beef cows (4 to 7 yr of age), feedlot steers (18 mo of age), free-ranging cow elk (3 to 5 yr of age), and chicken breast. Lipids were analyzed by capillary GLC. Total saturated fatty acids (SFA) were greater (P < 0.01) in range bison than in feedlot bison and were greater (P < 0.01) in SS of range beef cattle than in feedlot steers. Muscles of elk and range bison were similar (P > 0.05) in SAT. In LD, polyunsaturated fatty acids (PUFA) were highest (P < 0.01) for elk and range bison and lowest (P < 0.01) for feedlot steers within each muscle. Range bison and range beef cows had greater (P < 0.01) PUFA in LD and ST than feedlot bison or steers, respectively. Range-fed animals had higher (P < 0.01) n-3 fatty acids than feedlot-fed animals or chicken breast. Chicken breast n-6 fatty acids were greater (P < 0.01) than for muscles from bison, beef, or elk. Elk had higher (P < 0.01) n-6 fatty acids than bison or beef cattle; however, range-fed animals had higher (P < 0.01) n-6 fatty acids than feedlot-fed animals in ST. Conjugated linoleic acid (CLA, 18:2cis-9, trans-11) in LD was greatest (P < 0.01) for range beef cows (0.4%), and lowest for chicken breast and elk (mean = 0.1%). In ST, CLA was greatest (P < 0.01) for range and feedlot bison and range beef cows (mean = 0.4%) and lowest for elk and chicken breast (mean = 0.1%). Also, SS CLA was greatest (P < 0.01) for range beef cows (0.5%) and lowest for chicken breast (0.1%). Mean total fatty acid concentration (g/100 g tissue) for all muscles was highest (P < 0.01) for feedlot bison and feedlot cattle and lowest (P < 0.01) for range bison, range beef cows, elk, and chicken. Chicken breast cholesterol (mg/100 g tissue) was higher (P < 0.01) than LD and ST cholesterol, which were lowest (P < 0.01; 43.8) for range bison and intermediate for the other species. Cholesterol in SS was highest (P < 0.01) for feedlot bison and steers, which were similar to chicken breast (mean = 61.2 vs 52.8 for the mean of the other species). We conclude that lipid composition of bison muscle varies with feeding regimen, and range-fed bison had muscle lipid composition similar to that of forage-fed beef cows and wild elk.

Reduced asthma symptoms with n-3 fatty acid ingestion are related to 5-series leukotriene production.

Asthma may respond to dietary modification, thereby reducing the need for pharmacologic agents. This study determined the effectiveness of n-3 polyunsaturated fatty acid (PUFA) ingestion in ameliorating methacholine-induced respiratory distress in an asthmatic population. The ability of urinary leukotriene excretion to predict efficacy of n-3 PUFA ingestion was assessed. After n-3 PUFAs in ratios to n-6 PUFAs of 0.1:1 and 0.5:1 were ingested sequentially for 1 mo each; patient respiratory indexes were assessed after each treatment. Forced vital capacity (FVC), forced expiratory volume for 1 s (FEV1), peak expiratory flow (PEF), and forced expiratory flow 25-75% (FEF 25-75) were measured along with weekly 24-h urinary leukotriene concentrations. With low n-3 PUFA ingestion, methacholine-induced respiratory distress increased. With high n-3 PUFA ingestion, alterations in urinary 5-series leukotriene excretion predicted treatment efficacy. Elevated n-3 PUFA ingestion resulted in a positive methacholine bronchoprovocation dose change in > 40% of the test subjects (responders). The provocative dose to cause a 20% reduction (PD20) in FEV1, FVC, PEF, and FEF25-75 values could not be calculated because of a lack of significant respiratory reduction. Conversely, elevated n-3 PUFA ingestion caused some of the patients (nonresponders) to further lose respiratory capacity. Five-series leukotriene excretion with high n-3 PUFA ingestion was significantly greater for responders than for nonresponders. A urinary ratio of 4-series to 5-series leukotrienes < 1, induced by n-3 PUFA ingestion, may predict respiratory benefit.

Characterization of leukotriene production in vivo and in vitro in resident and elicited peritoneal macrophages in chickens and mice.

Previously, we reported differences in arachidonic acid metabolism in elicited chicken peritoneal macrophages when compared with murine resident and elicited peritoneal macrophages. We now describe leukotriene (LT) production in the same systems, using resident (murine) and inflammatory macrophages (from both species). Inflammatory (4- or 42-g Sephadex-elicited) peritoneal macrophages from chickens lacked the capacity to produce LT in vivo (following opsonized zymosan [OZ] stimulation) or in vitro, in response to A23187. In addition, chicken macrophages were unable to metabolize exogenously added LTC4 or LTD4 in vitro. In contrast, resident murine peritoneal macrophages produced measurable quantities of LTs (in vivo) within 5 min with an 8-fold increase after 45 min. LTC4 was effectively converted to LTE4 in vivo in a time-dependent manner (65% LTC4/35% LTE4 after 5 min stimulation with OZ and 6% LTC4/94% LTE4 after 60 min stimulation), but no in vitro. The lack of LTC4 metabolism to LTE4 in vitro could not be explained by cell-cell interaction between adherent and nonadherent cells. LTD4 was not detected under any experimental condition. Murine peritoneal cells incubated with LTD4 (with or without agonist) produced LTE4 in a time-dependent fashion. Addition of L-cysteine (a dipeptidase inhibitor) did not explain the lack of detectable levels of LTD4 following intraperitoneal stimulation with OZ. These results suggest that elicited chicken peritoneal macrophages are incapable of producing LTs compared to murine peritoneal macrophages. In addition, these studies fail to explain the different product profiles with in vivo stimulation of murine peritoneal macrophages as compared to in vitro stimulation.

Ingestion of n-3 polyunsaturated fatty acids and ovulation in rats.

The effects of different ratios of dietary (n-3):(n-6) polyunsaturated fatty acids on prostaglandin E, prostaglandin F2 alpha and ovulation in rats were assessed. Dietary (n-3) polyunsaturated fatty acids were incorporated, by ovarian phospholipids with ovarian tissue enrichment, with (n-3) polyunsaturated fatty acids enhancing, and (n-6) polyunsaturated fatty acids reducing, the number of ova released in immature rats primed with pregnant mares' serum gonadotrophin and human chorionic gonadotrophin. Incorporation of (n-3) polyunsaturated fatty acids appeared to enhance ovulation by altering total prostaglandin E production. This effect may be induced by changes in the prostaglandin E3:E2 ratio and the synthesis of less biologically active prostaglandin E3, or by dilution of the anti-ovulatory properties attributable to prostaglandin E2. High incorporation of dietary (n-6) polyunsaturated fatty acids may lead to reduced ovulation through excessive production of prostaglandin E2. Prostaglandin E or F2 alpha and alterations in tissue phospholipid composition inhibited progesterone release, and inhibition was independent of the series of prostaglandin produced. This study provides evidence that dietary lipids affect ovulation in rats with possible implications for reproduction in other vertebrates.

Caffeine-induced hypercalciuria and renal prostaglandins: effect of aspirin and n-3 polyunsaturated fatty acids.

Effects of aspirin and fish-oil ingestion on caffeine-induced hypercalciuria and renal prostaglandins (PG) were investigated in 12 healthy women. The 11-wk study consisted of 7-d baseline, 5-d aspirin (1000 mg/d), 11-d washout, and two 24-d fish-oil periods (FO-1 and FO-2, respectively, providing 1.5 vs 3.0 g n-3 fatty acids/d) separated by a 4-d washout period. Caffeine-load (CL) tests providing 5 mg caffeine/kg body wt were administered after baseline and each experimental period. Timed urine samples were collected precaffeine (basal) and at 1, 2, and 3 h post-caffeine. PGE2 excretion decreased during tests after aspirin and FO-2. There were significant increases in PGF2 alpha from baseline during each CL test. Hypercalciuria was observed during each CL test and the magnitude of this response was not altered by the experimental treatments. The finding that concentrations of post-caffeine urinary PGF2 alpha paralleled concentrations of urinary calcium supports the possibility that this prostaglandin plays a role in caffeine-induced hypercalciuria.

Frequency of (n-3) polyunsaturated fatty acid consumption induces alterations in tissue lipid composition and eicosanoid synthesis in CD-1 mice.

This study assessed the effect of frequency of consumption of a diet containing 10 g fat/100 g, with a (n-3):(n-6) polyunsaturated fatty acid (PUFA) ratio of 0.5, on tissue fatty acid composition and eicosanoid synthesis in CD-1 mice. With greater (n-3) PUFA consumption frequency, hepatic and peritoneal cell (n-3) PUFA levels changed to 14.4 +/- 1.3 and 12.4 +/- 0.9 mol/100 mol, respectively, from 2.4 +/- 0.3 and 1.9 +/- 0.3 mol/100 mol in control animals consuming a diet without (n-3) PUFA. Hepatic and peritoneal cell (n-6) PUFA levels were reduced to 6.4 +/- 0.5 and 7.6 +/- 0.5 mol/100 mol, respectively, with daily (n-3) PUFA consumption, compared with 23.6 +/- 0.7 and 17.9 +/- 0.7 mol/100 mol in control animals. Prostaglandin E, 6-keto-prostaglandin F1 alpha, and 4-series sulfidopeptide leukotriene biosyntheses reflected frequency of (n-3) PUFA consumption, with daily (n-3) PUFA consumption reducing prostaglandin E, consumption of (n-3) PUFA every other day reducing 6-keto-prostaglandin F1 alpha, and (n-3) PUFA consumption every 3 d reducing leukotriene C4 and leukotriene E4. Leukotriene C5 and leukotriene E5 changed from undetectable in animals consuming the control diet to > 40% of total leukotriene production in animals consuming (n-3) PUFA daily. Daily to biweekly consumption of fish or fish oil [1.26-0.63 g/wk of (n-3) PUFA, 6-3% of energy] may be a means for changing eicosanoid production. Consumption of (n-3) PUFA less often than twice a week may not change eicosanoid synthesis.

Evidence for mechanisms of the hypotriglyceridemic effect of n-3 polyunsaturated fatty acids.

Ingestion of modest amounts of n-3 polyunsaturated fatty acids (PUFA) (2.8% w/w) decreased plasma triglyceride levels in Syrian hamsters by 49%. This was associated with a 45% increase in hepatic carnitine palmitoyl transferase activity. Significantly, at this low level of n-3 PUFA intake, hepatic peroxisomal oxidation measured as CN- insensitive palmitoyl-CoA dependent NAD reduction was unaffected. Consumption of increasing amounts of dietary n-3 PUFA up to 2% (w/w) in hamster diets containing 15% fat, gradually decreased plasma triglycerides to 56% of the control levels. The diet induced changes in plasma triglyceride levels were highly correlated (r = -0.97, P less than 0.01) with changes in hepatic carnitine palmitoyl transferase activity. A gradual decrease up to 59% in hepatic phosphatidate phosphohydrolase activity with n-3 fatty acid consumption was also observed. The hypotriglyceridemic effects of moderate intakes of n-3 fatty acids are, therefore, associated with changes in key enzymes in hepatic triglyceride synthesis and mitochondrial oxidation, but not peroxisomal oxidation.

Dependence on dietary cholesterol for n-3 polyunsaturated fatty acid-induced changes in plasma cholesterol in the Syrian hamster.

Male Syrian hamsters consumed diets containing incremental increases in dietary n-3 fatty acids from fish oil with either low (0.015% w/w) or moderate (0.1% w/w) dietary cholesterol content. Animals consuming diets containing moderate cholesterol, but not animals consuming diets containing low cholesterol, had increased plasma very low (VLDL)- and low density lipoprotein (LDL)-cholesterol levels with increasing fish oil consumption. The plasma concentration of high density lipoprotein (HDL)-cholesterol decreased by 43 and 32% with the consumption of the highest fish oil diets in the low and moderate dietary cholesterol groups, respectively. Hepatic LDL-receptor binding activity did not change with the consumption of low cholesterol diets, but gradually decreased with fish oil consumption in animals consuming the moderate cholesterol diets. Hepatic LDL-receptor binding and plasma LDL-cholesterol levels of the different dietary fish oil groups were highly correlated (r = -0.91). Fish oil consumption also caused an increase in hepatic free cholesterol but a decreased cholesteryl ester content. Therefore, in the Syrian hamster, the consumption of n-3 fatty acids increases LDL-cholesterol levels which can be partially explained by decreased hepatic LDL-receptor binding and this response to dietary n-3 fatty acids is dependent on the dietary cholesterol content. However, the effects of dietary n-3 fatty acids on HDL-cholesterol are independent of dietary cholesterol content.

Dietary arachidonic and linoleic acids: comparative effects on tissue lipids.

The effects of preformed dietary arachidonic acid (AA, 20:4n-6) on murine phospholipid fatty acid composition in tissues capable (liver) and incapable (peritoneal exudate cells, PEC) of desaturating and elongating linoleic acid (LA, 18:2n-6) to AA were investigated. The results were compared with those obtained on matched animals on LA diets by either substituting or supplementing dietary LA with AA. Modest amounts of AA ethyl ester (0.5 wt%) included in the diet significantly increased tissue phospholipid AA levels by 39% and 57% in the liver and in PEC, respectively. The changes were further enhanced when dietary LA and AA intakes were equivalent, i.e., 57% and 68% in liver and PEC, respectively. This enrichment was observed in all phospholipid classes analyzed, with the greatest impact on phosphatidylcholine. In addition, the doubling of dietary LA had little effect on tissue phospholipid AA levels. The data suggest that while the level of n-6 PUFA may have an important effect on tissue fatty acid composition, the type of n-6 PUFA in the diet could be of greater significance.

The comparative effects of dietary alpha-linolenic acid and fish oil on 4- and 5-series leukotriene formation in vivo.

The comparative effects of dietary alpha-linolenic acid and fish oil on eicosanoid metabolism was studied in vivo. Resident murine peritoneal cells were stimulated in vivo with opsonized zymosan in animals maintained on diets containing increasing amounts of alpha-linolenic acid or fish oil concentrate with projected n-3/n-6 ratios of 0.2, 0.4 and 1.0. While fish oil feeding resulted in significant changes in eicosapentaenoate tissue levels, alpha-linolenic acid was preferentially metabolized to docosahexaenoate. High performance liquid chromatographic analysis revealed the formation of leukotriene E5 (LTE5) in all the fish oil groups (19.8 +/- 3.5 ng/mouse to 83.3 +/- 13 ng/mouse), but only in the highest linolenic acid group (6.0 +/- 3.2 ng/mouse). Concomitantly, the 4-series sulfidopeptide leukotrienes and PGI2 were significantly reduced in the two highest fish oil containing dietary groups. Similar reductions were observed in the highest linolenic acid group, but the changes were not statistically different from the control values. In summary, this paper reports the de novo synthesis of 5-series sulfidopeptide leukotrienes in animals consuming alpha-linolenic acid. It also reveals that dietary fish oil is 2.5 to 5 times more effective than alpha-linolenic acid in modulating eicosanoid metabolism and altering tissue phospholipid fatty acid composition.

Effect of increasing the dietary (n-3) to (n-6) polyunsaturated fatty acid ratio on murine liver and peritoneal cell fatty acids and eicosanoid formation.

An incremental increase in the dietary (n-3):(n-6) polyunsaturated fatty acid (PUFA) ratio from 0 to 1.93 in diets containing 15% fat (wt/wt) decreased the total (n-6) PUFA content of phospholipids of the liver and peritoneal cells (macrophage) in mice from 43.1 and 33.6 mol/100 mol to 16.0 and 12.3 mol/100 mol with a concomitant increase of 27.6 and 16.1 mol/100 mol in (n-3) PUFA, respectively. Consumption of (n-3) PUFA increased hepatic (n-3) PUFA levels without changing total PUFA (46.35 vs. 46.87 mol/100 mol), whereas macrophage PUFA levels were decreased. The synthesis of sulfidopeptide leukotrienes (SP-LT) (LTC4 and LTE4) was progressively reduced by increasing dietary (n-3) PUFA, i.e., there was a reduction of 76% in mice fed a diet containing a (n-3):(n-6) PUFA ratio of 1.93 compared with the control diet. The 5-series SP-LT (LTC5 and LTF5) were produced in all animals consuming (n-3) PUFA and accounted for 62% of all SP-LT synthesized in mice fed the diet containing a 1.93 (n-3):(n-6) PUFA ratio. Synthesis of 6-keto-prostaglandin F1 alpha decreased 81% in mice fed a diet containing a (n-3):(n-6) PUFA ratio of 1.93 whereas prostaglandin E2 synthesis decreased 44% in mice fed diets with (n-3):(n-6) ratios ranging from 0.41 to 1.93.

In vivo formation of leukotriene E5 by murine peritoneal cells.

Resident mouse peritoneal cells, stimulated in vivo with opsonized zymosan, produced leukotriene C4 and E4, with LTE4 being the major (80-90%) product. When mice were placed on diets containing increasing amounts of fish oil, four additional sulfidopeptide leukotrienes (SP-LT), LTC5, LTE5, 11-trans LTC5 and 11-trans LTE5, were identified. The identity of LTE5 was confirmed by spectrophotometric, chromatographic and enzymatic methods. When equivalent amounts of n-6 and n-3 polyunsaturated fatty acids (PUFA) were included in the diet, the stimulated peritoneal cells (in vivo) produced higher quantities of LTE5 (30.2 +/- 5.4 ng/10(6) cells) than LTE4 (22.8 +/- 7.3 ng/10(6) cells). In addition, in vitro studies demonstrated a 60% reduction in LTC4 (42.0 +/- 10.8 ng/10(6) cells to 16.7 +/- 6.2 ng/10(6) cells) and the appearance of LTC5 (2.1 +/- 0.9 ng/10(6) cells) in resident macrophages (stimulated with A23187) from mice maintained on a fish oil diet compared to mice fed the control diet. This study demonstrated that formation of the pentaenyl SP-LT in vivo, in particular LTE5, by peritoneal cells can significantly contribute to the endogenous SP-LT pool in response to an inflammatory stimulus following a dietary regimen containing fish oil.

Dietary unsaturated fatty acids: interactions and possible needs in relation to eicosanoid synthesis.

In addition to providing energy and essential fatty acids, dietary fatty acids can affect numerous biochemical and physiologic reactions related to secretory, cardiovascular, and immune functions. The major dietary unsaturated fatty acid, linoleic acid, affects tissue arachidonic acid and can influence eicosanoid-mediated reactions. Chronic, excess, or imbalanced eicosanoid synthesis may be conductive to excessive inflammation, thrombotic tendencies, atherosclerosis, and immune suppression. Dietary n-3 polyunsaturated fatty acids (PUFAs) may ameliorate eicosanoid-related phenomena by reducing tissue arachidonic acid and by inhibiting eicosanoid synthesis. This review summarizes information concerning the metabolism of unsaturated fatty acids, with emphasis on tissue arachidonic acid levels and eicosanoids, and discusses the need for data concerning the appropriate intake of dietary n-6 and n-3 PUFAs to modulate arachidonic acid and eicosanoid synthesis and to minimize possible adverse reactions.