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Biomedical subjects

A B Awad

Publications and source records attributed to A B Awad.

At least 37 records · Page 2Linked to original sources

In vitro activation and inhibition of rat colonic phospholipase D by fatty acids.

The present study was designed to examine the effect of long-chain fatty acids on phospholipase D (PLD) of the proximal colon mucosa of the rat. These fatty acids are present in the colon, in close contact with the membranes which contain the enzyme. The results indicate that unsaturated fatty acids activate the enzyme. At 4 mM, trienes are more potent activators as compared to 18:2 and 18:1. The number of double bonds and the configuration around the double bond were found to be important factors in activation of the enzyme. The metabolism of polyunsaturated fatty acids may be involved in the activation since the presence of antioxidants and cyclooxygenase inhibitors were found to influence the activation of PLD by polyunsaturated fatty acids.

Animals↗

Influence of dietary fat and feeding period on phosphoinositide metabolism in rat colonocytes.

The objective of the present study was to examine the effect of dietary fat content on phosphoinositide (PI) metabolism, fatty acid composition in colonocytes, and colonic luminal content of bile acids (BA) and free fatty acids (FFA) in rats. Male Sprague-Dawley rats weighing approximately 166 g were fed to semipurified diet containing 3% or 21.5% beef fat and 2% corn oil. The nonfat ingredients were adjusted to correct for differences in food consumption of these diets. Animals were fed these diets ad libitum for one or four weeks. The isolated colonocytes had a viability of 88.9% in all groups. PI metabolism was examined in the absence (basal) or presence of agonists, 2 mM deoxycholic acid or 10 microM A23187. Dietary fat concentration had no effect on PI metabolism, but the length of feeding had a significant effect on basal and stimulated PI metabolism. Colonocytes of animals fed the diets for four weeks were less sensitive to stimulation of PI cycle by agonists than those of animals fed for one week. Colonocyte fatty acid composition was influenced by dietary fat and feeding period. Only the relative percentage of 20:3(n-6) was significantly lower in rats fed the high-fat diet for one week; 18:0 was lower and 18:3(n-6) was higher in colonocytes of animals fed the diets for one week than in those fed for four weeks. Several colonic fatty acids, namely, 16:0, 20:3(n-6), and 22:5(n-6), also exhibited diet-by-feeding period interaction. Intracolonic luminal contents from rats fed the high-fat diet contained elevated concentrations of BA and FFA (44% and 62%, respectively). It was concluded that despite the effects of dietary fat concentration on increased colonic BA and FFA and on altered membrane fatty acid composition, dietary fat had no effect on PI metabolism in colonocytes under the conditions in the present experiment. A difference in components between the purified diet and the commercial rat chow and/or an aging effect of the rats may alter the PI cycle of colonocytes.

Animals↗

Differential response of glucose utilization by three rat muscle tissues to dietary fatty acid composition.

Rats were fed a semisynthetic diet containing 14% of either beef fat, safflower oil, or menhaden oil plus 2% corn oil for 7 weeks, and three tissues, diaphragm, heart, and skeletal muscle, were examined for fatty acid composition in their phospholipids and triglycerides. In addition, the lipid concentrations in these tissues were examined. The in vitro oxidation and incorporation of glucose into lipids of these tissues were also examined. Skeletal muscle showed the greatest change in phospholipid composition with diet. All tissues were responsive to changes in diet in regards to the triglyceride fraction. Dietary alteration of tissue phospholipid composition did not alter lipid concentration in these tissues. However, in diaphragm tissue, rats fed the beef fat diet had lower phospholipid and higher triglyceride synthesis compared with those fed either menhaden oil or safflower oil. In addition, triglyceride synthesis was higher in the diaphragm of animals fed the menhaden oil diet as compared with the safflower oil diet. Therefore, dietary fatty acid composition may play a role in the triglyceride and phospholipid metabolism of rat diaphragm.

Animals↗

Effect of dietary fat on metabolic adjustments to maximal VO2 and endurance in runners.

The present study examined the effects of dietary manipulations on six trained runners. The percent energy contributions from carbohydrate, fat, and protein were 61/24/14, 50/38/12, and 73/15/12 for the normal (N), fat (F), and carbohydrate (C) diets, respectively. Expiratory gases and blood responses to a maximum (VO2max) and a prolonged treadmill run were determined following 7 d on each diet. Free fatty acids (FFA), triglycerides, glycerol, glucose, and lactate were measured. Dietary assessment of subjects' N diet indicated that they were consuming approximately 700 kcal.d-1 less than estimated daily expenditures. Running time to exhaustion was greatest after the F diet (91.2 +/- 9.5 min, P < 0.05) as compared with the C (75.8 +/- 7.6 min, P < 0.05) and N (69.3 +/- 7.2 min, P < 0.05) diets. VO2max was also higher on the F diet (66.4 +/- 2.7 ml.kg-1 x min-1, P < 0.05) as compared with the C (59.6 +/- 2.8 ml.kg-1 x min-1, P < 0.05) and N (63.7 +/- 2.6 ml.kg-1 x min-1, P < 0.05) diets. Plasma FFA levels were higher (P < 0.05) and glycerol levels were lower (P < 0.05) during the F diet than during the C and N diets. Other biochemical measures did not differ significantly among diets. These data suggest that increased availability of FFA, consequent to the F diet, may provide for enhanced oxidative potential as evidenced by an increase in VO2max and running time. This implies that restriction of dietary fat may be detrimental to endurance performance.

Analysis of Variance↗

Alteration of membrane fatty acid composition and inositol phosphate metabolism in HT-29 human colon cancer cells.

The present study was designed to investigate the role of membrane fatty acid (FA) composition on inositol phosphate (InsP) release by a human colon tumor cell line. Cells were supplemented for five days in culture with 0, 10, 30, or 100 microM sodium stearate (18:0), linoleate [18:2(omega-6)], or linolineate [18:3(omega-3)]. These FAs were supplied as a complex with FA-free bovine serum albumin. InsP release was examined in these cells with or without stimulation with deoxycholic acid (DCA) after they were labeled with [3H]myoinositol. FA enrichment was found to influence inositol incorporation into membrane lipids. Although 18:0 had no effect, 18:2(omega-6) decreased the incorporation. On the other hand, 18:3(omega-3) increased the incorporation of inositol compared with the cells supplemented with the other FAs, but they were not different from control. Basal release of total InsP was elevated only with supplementation of 10 and 30 microM 18:3(omega-3). FA supplementation with 18:0 at 30 microM and 18:2 at 30 and 100 microM resulted in downregulation of bsal release of InsP. Enrichment of HT-29 cell membranes with polyunsaturated FAs resulted in a significant increase in stimulated release of InsP, but this was not seen with saturated FA supplementation. At 10 microM supplementation, 18:2 had the greatest effect on stimulated InsP release. This effect of 18:2 disappeared at 30 microM. However, the increase in the stimulated InsP release caused by 18:3 occurred at 10 and 30 microM. DCA-stimulated release of InsP was not downregulated by any FA supplementation. This study showed that enrichment of the membranes with polyunsaturated FAs increases the response of the phosphatidylinositol cycle to DCA stimulation. In addition, enrichment with 18:3(omega-3) increases the basal turnover of InsP. It is concluded that alteration of membrane FAs has a profound effect on the phosphatidylinositol cycle.

Colonic Neoplasms↗

Differential effect of butyrate on lipids of human colon cancer cells.

Recently, we demonstrated that treatment of LS174T cells with 2 mM butyrate for one day had a significant effect on the composition of cellular fatty acids. In an attempt to further explore this phenomenon, we investigated the effect of long-term butyrate treatment in the presence of different fatty acids in the medium on cellular phospholipids (PLs) and triacylglycerol (TG). Cells were supplemented with 100 microM sodium salts of 18:2 (n-6), 20:4 (n-6), 20:5 (n-3), or 22:6 (n-3) as a fatty acid-free-albumin complex. The molar ratio of the albumin and these long-chain fatty acids (LCFAs) was 3:1. One-half of these cultures were supplied with 2 mM butyrate, and the pH was adjusted to 7.4. The supplementation of the LCFAs and butyrate was maintained for eight days. The present study indicates that butyrate had a differential effect on the fatty acid composition of PLs and TG of LS174T cells. This includes an increase in monounsaturates and elongation of the supplemented LCFA, and this effect was more pronounced on TG than PL fatty acids. Butyrate resulted in a significant reduction in polyunsaturated fatty acid concentration only in PLs. In general, butyrate decreased the unsaturation index (UI) of the PLs but increased that of TG. The present study also confirmed our previous observation regarding the effect of LCFAs on cellular lipids. PL and TG fatty acid chain lengths reflect those of supplemented fatty acids. The UI of these two lipid fractions increased more with supplementation of n-3 than n-6 fatty acids.(ABSTRACT TRUNCATED AT 250 WORDS)

Butyrates↗

Effect of dietary fat on glomerular lipid composition and angiotensin II receptors.

Nutritional factors are major etiologic determinants which may affect the incidence or severity of hypertension. Since angiotensin II (Ang II) has a role in some forms of hypertension, the influence of dietary lipid composition on glomerular lipid content and on Ang II receptor parameters was determined. Three groups of rats were fed diets containing 14% by weight of 1) beef fat (saturated fatty acid rich), 2) safflower oil (n-6 fatty acid rich), or 3) fish oil (n-3 fatty acid rich), each supplemented with 2% corn oil. A fourth group of rats was fed rat chow which contained 6.5% fat. At the end of 7 weeks, the rats were sacrificed, the kidneys removed, and renal glomeruli isolated using a sieving technique. Feeding diets which varied in the quantity and composition of fatty acids altered glomerular fatty acid composition of n-6 and n-3 fatty acids and the unsaturation indices of glomeruli. The cholesterol content, but not the phospholipid content, was decreased in glomeruli of animals fed the high fat diets. This resulted in a lower cholesterol/phospholipid ratio, indicative of an increase in membrane fluidity. Glomerular binding was determined using 125I-Ang II in a radioreceptor assay. Binding was proportional to glomerular protein and was time dependent. Receptor affinity (Ka) and number were determined by Scatchard analysis of binding inhibition data. Glomerular Ka was significantly lower for animals fed the rat chow as compared to the semisynthetic diets. There was no significant difference in the concentration of Ang II receptors in glomeruli of animals fed the different diets. The data indicate that the induced alteration in glomerular lipid composition is associated with alteration in the binding affinity of glomerular Ang II receptors.

Animals↗

Influence of butyrate on lipid metabolism, survival, and differentiation of colon cancer cells.

The present work was designed to study the differentiating effect of butyrate on LS174T cells after modification of their lipids with long-chain fatty acid (LCFA) supplementation. The LCFAs 18:1(n-9), 18:2(n-6), 20:4(n-6), 20:5(n-3), and 22:6(n-3) bound to added to the media of confluent cells for eight days. The fatty acid-to-albumin ratio was 3:1. The concentration of fatty acids in the media was 100 microM. On the last day, half of the flasks were treated with 2 mM butyrate. The data indicate that supplementation with polyunsaturated LCFAs having 20-22 carbon atoms resulted in a significant reduction in cell density and viability, whereas all LCFA supplementation reduced differentiation as measured by alkaline phosphatase activity. Butyrate treatment increased the density, viability, and differentiation of the tumor cells. The effect of butyrate on differentiation was mainly with cells supplemented with 18:1, 20:5, and 22:6. In the absence of LCFA supplementation, butyrate reduced the concentration of 22:5(n-6) in the cellular lipids. Also, butyrate modified the LCFAs incorporated in cells supplemented with 18:2 and 20:5, with changes occurring in 20:5(n-3), 22:5(n-3), and 22:5(n-6). Thus the present study suggests an interaction between butyrate and LCFA on differentiation and LCFA metabolism of human colon cancer cells.

Adenocarcinoma↗

Dietary fat and phospholipase A2 activity of Sprague-Dawley rat large intestine.

The present studies were conducted to examine the effect of dietary lipid content and composition [(n-6) vs. (n-3) fatty acids] on the activity of mucosal phospholipase (PL)A2 of the large intestinal tract of rats. Three segments of the large intestinal tract were examined: cecum, proximal colon and distal colon. Weanling male Sprague-Dawley rats were fed diets containing either 5% (LS) or 16% safflower (HS) or 14% menhaden oil plus 2% safflower oil (HM) for 3 wk with the oil replacing starch in the HS and HM diets on a weight basis. The lipid extracts of microsomal fractions from mucosal scrapings were examined for phospholipid and cholesterol content and fatty acid composition. Phospholipase A2 was assayed using a fluorescent substrate. Rats fed the high fat diets had lower PLA2 specific activities. The (n-3) or (n-6) fatty acid enrichment of the membranes had no effect of the activity of the enzyme. The activity of the enzyme decreased aborally from the cecum to the distal colon; the proximal colon had an intermediate specific activity.

Animal Nutritional Physiological Phenomena↗

Effect of excess dietary calcium on colon mucosal membranes and fecal lipids.

The objective of the present studies was to examine the biochemical alterations in colon apical membranes upon feeding excess calcium to animals on a saturated fatty acid-rich diet. It has been suggested recently that excess dietary calcium may offer a protection to colon membranes as judged by histologic examination. Sprague-Dawley weanling male rats were fed a semisynthetic diet containing 14% beef fat plus 2% corn oil and either the calcium requirement or excess calcium in the form of calcium carbonate. Animals were fed the diets for 4 weeks. Feces were collected in the last 3 days. The results indicate that excess dietary calcium resulted in alteration in the density of 4 protein bands of colon apical membranes upon examination on SDS-gel electrophoresis. These bands contain 20% of membrane proteins. The diet had no effect on either the lipid content or fatty acid composition of the membranes. Excess dietary calcium resulted in a 54% reduction in fecal water bile acids and a 44% reduction in fecal water free fatty acids. The reduction in fecal water lipids was due to alterations in the solubility of these lipids. This was not mediated through alterations in the pH of fecal water. The observed alterations in protein patterns of these membranes may be due to either the reduction of fecal water bile acids and free fatty acids or may be a direct effect of dietary calcium on membrane proteins.

Animals↗

Effect of dietary fat on the lipid composition and utilization of short-chain fatty acids by rat colonocytes.

The objective of the present studies was to examine the effect of dietary fat on the lipid composition of rat colonocytes and their utilization of short-chain fatty acids (SCFA). Rats were fed 14% beef fat, fish oil or safflower oil plus 2% corn oil in a semi-synthetic base diet for 4 wk. Colonocytes were isolated and their lipid composition was examined. Feeding beef fat and fish oil resulted in an increase in monounsaturated fatty acids and a reduction in omega-6 fatty acids. Feeding fish oil resulted in an enrichment with omega-3 fatty acids. There was no dietary influence on the amount of either cholesterol or phospholipids of colonocytes. Fish oil feeding resulted in significant increase in colonocyte free fatty acids (FFA) as compared to other diets. Dietary fat was found to have no effect on SCFA utilization by colonocytes. Colonocytes were found to utilize SCFA in the order of butyrate greater than or equal to acetate greater than or equal to propionate. The presence of acetate and propionate in the medium had no effect on the rate of butyrate utilization.

Animals↗

Failure to demonstrate an effect of dietary fatty acid composition on body weight, body composition and parameters of lipid metabolism in mature rats.

The objectives of the present study were to examine the effects of dietary fatty acid composition on body composition and on several parameters of lipid metabolism in adipose tissue and to assess the ease of weight loss with restricted dietary intake. Rats were fed diets containing 14% fish oil, safflower oil or beef fat plus 2% corn oil. These diets were fed ad libitum for 4 wk (first phase). A number of the animals from each group were killed, and the others were fed 50% of their first-phase food intake for an additional 4 wk (second phase). The diets used in the second phase contained 3% of the above fats plus 2% corn oil. Food consumption, food efficiency and body weights were monitored. After the rats were killed, fatty acid composition of epididymal fat pads was examined. In addition, in vivo lipolysis and in vitro lipogenesis in epididymal fat pads were examined. The results indicate that dietary fatty acid composition had no effect on body weight, food consumption, in vivo lipolysis and in vitro lipogenesis in epididymal fat pads. In addition, although dietary fatty acid manipulation resulted in alteration in adipose tissue fatty acid composition, it had no effect on the rate of weight loss, body composition, in vivo lipolysis and in vitro lipogenesis in epididymal fat pads. It is concluded that dietary fatty acid composition does not play a role in body composition and in lipid metabolism in adipose tissue of mature rats.

Animals↗

Effect of dietary fat composition on rat colon plasma membranes and fecal lipids.

The present study was designed to examine the effect of dietary fat composition on the structure of colon mucosal plasma membranes and fecal lipids. Rats were fed a purified diet containing 14% of either highly saturated fat (beef fat or butterfat) or highly polyunsaturated oil (safflower) in addition to 2% corn oil for 4 wk. Colon mucosal membranes were prepared and examined for lipid composition and protein pattern. Saturated fatty acid feeding resulted in the loss of some protein bands from plasma membranes compared to feeding polyunsaturated fatty acids. Within the saturated fatty acid--rich fats, feeding beef fat caused a greater loss than did feeding butterfat. Dietary fat composition had no effect on membrane content of phospholipid and cholesterol. Saturated fatty acid feeding resulted in an increase in the percentage of 18:1 in plasma membrane lipids compared to feeding safflower oil. The observed changes in the structure of colon mucosal membrane of animals fed the saturated fats were associated with an increase in fecal free fatty acids. There was a 4-fold and 2-fold increase in fecal free fatty acids with feeding the beef fat and butterfat diets, respectively, compared to the safflower oil diet. Alterations in fecal bile acid and free fatty acid composition were also noticed with feeding saturated fatty acids. The results obtained suggest that feeding saturated fatty acids as the main source of fat in the diet could influence the structure of colon mucosa, and this could be mediated through fecal free fatty acids.

Animal Feed↗

Effect of dietary saturated fatty acids on hormone-sensitive lipolysis in rat adipocytes.

The objective of this work was to examine the mechanism by which dietary saturated fatty acids, as compared with polyunsaturated fatty acids, lower hormone-sensitive lipolysis in rat adipocytes. Rats were fed a purified diet containing 14% of a fat with a high concentration of either saturated fatty acids (coconut oil or beef fat) or polyunsaturated fatty acids (safflower oil) as a control. In addition, each diet contained 2% corn oil. The animals were fed these diets for 4 wk. Norepinephrine-stimulated lipolysis was 50% lower when diets rich in saturated fatty acids, regardless of their chain length, were fed than when a diet containing a high concentration of polyunsaturated fatty acids was fed. The specific activities of adenylate cyclase, 3',5'-cyclic nucleotide (cAMP) phosphodiesterase and hormone-sensitive lipase were lower when saturated fatty acids were fed than when polyunsaturated fatty acids were fed. Accumulation of cAMP upon stimulation with 10(-5) M norepinephrine was lower when saturated fatty acids were fed than when polyunsaturated fatty acids were fed. Moreover, adipocytes were larger when saturated fatty acids were fed than when polyunsaturated fatty acids were fed. The data obtained suggest that dietary saturated fats exert their inhibitory effect on hormone-stimulated lipolysis by influencing several points in the lipolytic cascade.

3',5'-Cyclic-AMP Phosphodiesterases↗

Effect of dietary saturated fatty acids on intracellular free fatty acids and kinetic properties of hormone-sensitive lipase of rat adipocytes.

Experiments were conducted to examine the effects of dietary saturated fatty acids on the intracellular free fatty acid concentration and composition and on the kinetic parameters of hormone-sensitive lipase of rat adipocytes. Animals were fed for 4 wk 14% coconut oil, beef fat or safflower oil and 2% corn oil in a purified diet. Adipocytes of animals fed the coconut oil diet contained higher basal level of intracellular free fatty acids than those of animals fed other beef fat or safflower oil diets. Norepinephrine (10(-5) M) stimulated the basal intracellular free fatty acid concentration by 2.3-3.4-fold in adipocytes from animals fed saturated fatty acids, compared with 6.4-fold in those of animals fed the safflower oil diet. The concentrations of intracellular free fatty acids in adipocytes of experimental animals after stimulation with 10(-5) M norepinephrine, however, were not significantly different. The intracellular free fatty acid pool of adipocytes of animals fed the saturated fatty acids had more palmitic acid and less linoleic acid than those of safflower oil-fed animals. The results indicate that type of dietary fat had no effect on kinetic properties of hormone-sensitive lipase.

Adipose Tissue↗

Developmental alterations in 5'-nucleotidase kinetics and lipid composition of rat heart sarcolemma.

The objective of these studies was to examine the mechanism by which the specific activity of heart sarcolemma 5'-nucleotidase decreases as function of age. We examined the kinetic properties and the lipid composition of the sarcolemma from animals with different ages. The age groups used were 1 month, 6-8 months and 13-15 months. It was found that the Km of this enzyme increases as the animal develops from 1 month to 6-15 months. The opposite was true with 5'-nucleotidase Vmax. There was no significant difference between the middle age and the older age groups in those parameters. The results of these experiments suggest that the increase in Km in sarcolemma 5'-nucleotidase could be due to the reduction of the sarcolemmal polyunsaturated fatty acid concentration, the only lipid alteration observed.

5'-Nucleotidase↗

Alteration of rat heart sarcolemma lipid composition by dietary elaidic acid.

Male weanling rats were fed a semisynthetic diet containing 5% of either elaidic acid or oleic acid for 4 weeks. The heart sarcolemma were isolated and examined for purity by using marker enzymes. The sarcolemma preparations were enriched in phosphodiesterase I, 5'-nucleotidase and p-nitrophenyl phosphatase as compared with the heart homogenates. Succinic dehydrogenase activity was not detected in these preparations. The lipids of sarcolemma fractions from animals fed the trans fatty acids were enriched with the fatty acid fed. Most of this incorporation was found to be associated with the two major phospholipids, phosphatidylcholine and phosphatidylethanolamine. The incorporation of elaidic acid into the membranes resulted in an increase in membrane cholesterol without affecting either membrane phospholipid content or profile. The alterations induced in the lipid composition of heart sarcolemma by feeding this dietary trans fatty acid suggest some functional alterations in these membranes.

Animals↗