James M. Iacono

Dietary α-linolenic acid alters tissue fatty acid composition, but not blood lipids, lipoproteins or coagulation status in humans

We examined the effect of dietary α-linolenic acid (ALA) on the indices of lipid and coagulation status and on the fatty acid composition of serum and peripheral blood mononuclear cell (PBMNC) lipids in ten healthy men (age 21–37 yr) who consumed all their meals at the Western Human Nutrition Research Center for 126 d. There was a stabilization period of 14 d at the start when all 10 subjects consumed the basal diet (BD) containing 23.4 energy percent (en%) fat and two intervention periods of 56 d each. During the first intervention period, 5 subjects consumed the BD containing 23.4 en% fat, and 5 subjects consumed a diet providing 6.3% calories from α-linolenic acid [flaxseed oil (FSO) diet containing 28.8 en% fat]. Diets were crossed over between the two groups during the second intervention period. Feeding the FSO diet did not nignificantly alter serum triglycerides, cholesterol, highdensity lipoproteins, low-density lipoproteins, apoprotein A-I and apoprotein B when compared to the corresponding values in the subjects fed the BD, nor was there any effect of the FSO diet on the bleeding time, prothrombin time and partial prothrombin time for these subjects. Feeding the ALA-containing diet did cause a significant increase in ALA concentration in serum (P<0.001) and PBMNC lipids (P<0.05). It also caused a significant increase (P<0.05) in the eicosapentaenoic and docosapentaenoic acid contents of PBMNC lipids, and a decrease (P<0.01) in linoleic and eicosatrienoic acid contents of serum lipids. Thus, dietary ALA, fed for 56 d at 6.3% of calories, had no effect on plasma triglyceride or very low density lipoprotein levels or the common risk factors associated with atherosclerosis, although these parameters have been reported by others to be influenced by fatty acids, such as palmitic or linoleic acids, in the diet. Dietary ALA did significantly alter the fatty acid composition of plasma and PBMNC.

Reduction in blood pressure associated with high polyunsaturated fat diets that reduce blood cholesterol in man

Abstract Ten male and 11 female subjects in the 40–60 age group were fed a 25% fat-calorie diet (Period I) with a P/S ratio of 1. The diet consisted of foods commonly eaten in the U.S.A. and was fed for 40 days, followed by a second 40-day feeding period of 35% fat calories (Period II) with a P/S ratio of 1. A significant decrease in systolic pressure for males and females was observed at the end of the 25% fat-calorie period. The lowered levels of systolic pressure were maintained throughout the 35% fat-calorie period. The diastolic pressure also decreased significantly during Period I; it showed a slight increase, however, during Period II. Plasma cholesterol was also significantly decreased during the first phase of the study and remained lowered during the second phase. Beta cholesterol response paralleled that of total cholesterol, while alpha cholesterol response paralleled that of percentage alpha lipoprotein lipid. Plasma lipoproteins reflected the changes in diet; percentage of pre-beta lipoprotein lipid rose during Period I, while percentage of alpha lipoprotein lipid fell. These results were reversed during Period II. No significant changes in percentage of beta lipoprotein lipid were observed. Blood urea nitrogen was significantly decreased during Period I, followed by a rise after Period II. Blood glucose tended to fall during Period II but rose above prestudy levels after the study. No significant changes were observed in body weights or plasma triglycerides. These results suggest that dietary fat may be one more important factor in elevated blood pressure in otherwise healthy subjects, and its influence should be investigated further in a larger controlled trial with both normo- and hypertensive subjects.

Concentration of dietary n-6 polyunsaturated fatty acids and the human immune status

We examined the effect of the dietary concentration of total fat and n-6 polyunsaturated fatty acids (PUFA) on the immune status of seven healthy women (age 30-65 years) who lived at our metabolic suite. During the first 20 days all subjects consumed a stabilization diet that contained 5.2 energy percent (en%) PUFA and 41.1 en% fat. For the next 40 days, three subjects consumed a diet with 3.2 en% PUFA and 26.1 en% fat, while the remaining four subjects consumed a diet with 9.1 en% PUFA and 31.1 en% fat. For the next 40 days, the diets of the two groups were crossed over. Blastogenesis of peripheral blood mononuclear cells cultured with phytohemagglutinin, concanavalin A, protein A, and pokeweed, and the serum concentrations of complement fractions C3 and C4 were significantly increased upon the feeding of both low fat (26.1 or 31.1 en%) diets compared to the values when the high fat (41.1 en%) diet was fed. None of the indices tested were different when the high PUFA (9.1 en%) and low PUFA (3.1 en%) diets were compared. Our results indicate that low fat diets improve some of the indices of human immune status and that a moderate increase in the level of n-6 PUFA in an otherwise low fat diet does not suppress the human immune system.

Dietary fat and blood pressure: an intervention study on the effects of a low-fat diet with two levels of polyunsaturated fat.

The role of dietary fat in human blood pressure control was studied among 84 middle-aged subjects (mainly couples) in two semirural communities in North Karelia, Finland. The families were randomly allocated into two groups that, after a baseline period of 2 weeks, changed their diet for a 12-week intervention period so that the proportion of energy derived from fats was similarly reduced in both groups, from 38 to 24%, but the polyunsaturated/saturated fatty acid (P/S) ratio was increased--from 0.2 to 0.9 in group I and to 0.4 in group II. After the intervention period, both groups switched back to their usual diet for a period of 5 weeks. During the intervention period, total serum cholesterol was reduced by 16% in group I and 14% in group II. Mean body weight and urinary sodium, potassium, calcium, and magnesium excretion changes were small or nonexistent. Mean systolic blood pressure decreased 4 mm Hg in group I (P less than 0.01) and 3 mm Hg in group II (P less than 0.01), and mean diastolic blood pressure decreased 5 mm Hg (P less than 0.001) and 4 mm Hg (P less than 0.01), respectively. The reductions were reversed during the switch-back period (P less than 0.01). These results confirm previous findings of the blood-pressure-reducing effect of a low-fat/high-P/S diet. Although a number of possible confounding factors can be ruled out, the dietary constituent accounting for the blood pressure change cannot be ascertained definitely. The results showed no significant further blood pressure reduction with more than a moderately increased P/S ratio when the saturated fat intake was markedly reduced.

Dual-labeled technique for human lipid metabolism studies using deuterated fatty acid isomers.

Two deuterated fatty acids, elaidate-d2 and oleate-d4, were fed simultaneously to a human subject as a mixture of trielaidin-d6 and triolein-d12. Periodically, blood samples were drawn, and red blood cells were separated from the plasma. Red blood cells and plasma lipids were fractionated and analyzed by combined gas chromatography—multiple ion mass spectroscopy. Dual deuterium-labeling allows rate and extent of fatty acid incorporation to be followed in various plasma and red cell neutral and phospholipid fractions. Maximum amount of deuterated fat varied from 4% in cholesterol ester to 64% in phosphatidyl ethanolamine. The highest levels of deuterated fat occurred in either 6-, 8-, or 12-hr samples; generally, <1% labeled fatty acids could be detected in 72-hr samples. Because the method is based on dual-labeling, differences in the relative incorporation of both fatty acid isomers can be compared directly. Differences in rates of incorporation, rates of removal, and extent of incorporation of labeled fatty acids into blood plasma can also be determined reliably. Our experimental labeling of fats with deuterium permits for the first time the metabolism of two fatty acid isomers to be compared simultaneously in human subjects. This new method should be applicable to a variety of other lipid metabolic studies.

Nutritional modulation of human immune status

Abstract Modulation of the immune status by a nutritionally balanced diet containing different levels of the polyunsaturated fatty acids (PUFA), with total fat porviding 25–30 energy % (en %), was examined in 8 healthy men (29–44 years) living in a metabolic unit. For the first two weeks all subjects were fed a stabilization diet (PUFA 6 en %), and for the next 11 weeks the level of PUFA calories was 3.5 en % for 4 subjects and 12.9 en % for the other 4 subjects. The level of PUFA in the diet did not affect any of the indices of the immune status tested. In both dietary groups, several indices of the immune status including the numbers of circulating T and B lymphocytes and their in vitro proliferation in response to T as well as B cell specific mitogens increased significantly with time. The numbers of circulating neutrophils and the serum concentration of complement fraction C3 decreased significantly in both the groups. Blood leukocyte count, serum contcentrations of IgA, IgE, IgG, IgM, and C4 remained unchanged in both groups. The immune status of these men was improved during the experimental period. Possible reasons for this improvement might include reduction in calories from fat or balanced and adequate supply of other nutrients.

Lack of effect on blood pressure by low fat diets with different fatty acid compositions

We compared the effects on blood pressure (BP) of three isocaloric diets with reduced total fat and saturated fatty acid (SAFA) contents but with different proportions of monounsaturated (MUFA) and polyunsaturated fatty acids (PUFA). Diet LF (low fat) provided 20 en% fat (7.9% SAFA, 7.8% MUFA, 3.0% PUFA); diet HP (high PUFA) 26 en% fat (7.5% SAFA, 8.2% MUFA, 8.1% PUFA), and diet HM (high MUFA) 26 en% fat (7.3% SAFA, 14.1% MUFA, 3.2% PUFA). The diets were consumed for 8 weeks (intervention) preceded by 2 weeks and followed by 8 weeks on a habitual diet (baseline/ switchback) with 33–34 en% fat (13–14% SAFA, 12% MUFA, 6% PUFA). Forty-five free-living couples were randomly allocated into the three diet groups, and 43 men and 44 women completed the study. BP was measured weekly with an automatic device. Compliance to diet was monitored by repeated food records, serum fatty acid compositions, and weekly visits to a nutritionist. Both systolic BP (SBP) and diastolic BP (DBP) remained unchanged throughout the study in all three groups. The weight-adjusted mean (s.e.m.) BP values showed changes in SBP of +1.7 (1.8), −0.4 (1.7), and +1.9 (1.9) mm Hg on the LF, HP, and HM diets, respectively (difference NS), and DBP of +0.1 (1.0), +0.6 (1.0), and −0.3 (1.0) mm Hg, respectively (difference NS) between the last 2 weeks of the baseline and intervention periods. The expected fatty acid intakes were achieved, and there were no between-group differences in change of body weight, intake of dietary fibre and potassium, and 24-h sodium excretion. A reduction in total fat and SAFA intake and changes in the proportions of dietary MUFA and PUFA did not affect the BP levels of this normotensive population with an adequate intake of PUFA at baseline.

Lability of red blood cell membranes to lipid peroxidation: Application to humans fed polyunsaturated lipids

Red blood cell membranes (RBCM) were used to estimate human red blood cell lability to lipid peroxidationin vitro. RBCM were prepared from blood collected from humans fed diets with either 3 or 15% polyunsaturated fatty acids for 80 days. RBCM were isolated by centrifugation, and oxidative stress was induced byin vitro incubation with 0.1 or 0.5 mM tert-butyl hydroperoxide (t-BOOH) in the presence of 0.5 mg added hemoglobin. Lipid Peroxidation was evaluated by measurement of thiobarbituric acid-reactive substances (TBARS). Lipid peroxidation correlated with the protein content of RBCM in both noninduced and t-BOOH-induced lipid peroxidation systems. TBARS production was dependent on the amount of t-BOOH added to the RBCM. The production of TBARS by RBCM incubated with 0.5 mM t-BOOH was correlated with the arachidonic acid content in the red blood cells (RBC) from which RBCM were prepared. The methodology developed was useful for comparative estimations of the lability of RBCM to lipid peroxidation.

The role of dietary polyunsaturated fatty acids and prostaglandins in reducing blood pressure and improving thrombogenic indices

Evidence linking dietary fats to blood pressure and thrombogenic indices is reviewed. Results of dietary studies performed at Beltsville, Maryland, have demonstrated that under controlled dietary conditions, i.e., when total fat intake is maintained at 25% fat calories with a P/S ratio of 1, at either a fixed or free-choice salt intake and where the body weight is maintained relatively constant, blood pressure can be lowered and platelet aggregation indices can be improved in men and women in the 40-60 age group. Results of a pilot epidemiologic study of farmers aged 40-45 in Finland and Italy generally confirm the experimental nutrition studies reported above. A possible explanation of these results based on the conversion of linoleic acid to prostaglandins as well as the physiological actions of prostaglandins is discussed.

Effects of reduced-fat diets with different fatty acid compositions on serum lipoprotein lipids and apolipoproteins.

OBJECTIVE To compare the effects on serum lipoproteins of three isocaloric diets with reduced total fat and saturated fatty acid (SFA) contents but with different proportions of monounsaturated (MUFA) and polyunsaturated fatty acids (PUFA). DESIGN A low-fat diet (LF) provided 20 en% fat, 7.9% SFA, 7.8% MUFA, 3.0% PUFA; a high-PUFA diet (HP) 26 en% fat, 7.5% SFA, 8.2% MUFA, 8.1% PUFA; and a high-MUFA diet (HM) 26 en% fat, 7.3% SFA, 14.1% MUFA, 3.2% PUFA. Diets were consumed for 8 weeks in a parallel design, after 2 weeks on a habitual diet with 33-34 en% fat, 13-14% SFA, 12% MUFA, 6% PUFA, and followed by an 8-week period on habitual diet. Compliance to diet was monitored by repeated food records and weekly visits to a nutritionist. SUBJECTS 45 free-living, middle-aged couples who were randomly allocated into the three diet groups; 43 men and 44 women completed the study. RESULTS During the diet periods, a small but significant reduction in body weight of 0.4-1.0 kg was observed in all groups. Similar and significant reductions of mean weight-adjusted serum total cholesterol (4-8%), low-density lipoprotein (LDL) cholesterol (7-11%), and high-density lipoprotein (HDL) cholesterol (8-11%) were observed during the diets. HDL2-cholesterol and apoprotein (apo) A-I levels were reduced whereas HDL3-cholesterol and apoA-II increased. ApoB was significantly decreased during the HM diet only. Serum triglycerides increased significantly during diets LF (25%, P < 0.01) and HP (19%, P < 0.05) but not during diet HM (5%, NS). CONCLUSIONS Reduction in the intake of total fat and saturated fatty acids reduced serum LDL-cholesterol and HDL2-cholesterol concentrations irrespective of the relative proportions of MUFA and PUFA in the diets. The results suggest that there might be some advantage in increasing the proportion of MUFA in low-fat diets, since the HM diet rich in MUFA reduced apoB and slightly attenuated the increase in serum triglycerides that is commonly associated with dietary fat reduction.